Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Motoyuki Kohima Last modified date:2021.07.29

Associate Professor / Department of Medicine and Bioregulatory Science, Graduate School of Medical Sciences, Kyushu University / Department of Clinical Medicine / Faculty of Medical Sciences


Papers
1. Kuwano A, Tanaka M, Suzuki H, Kurokawa M, Imoto K, Tashiro S, Goya T, Kohjima M, Kato M, Ogawa Y., Upregulated expression of hypoxia reactive genes in peripheral blood mononuclear cells from chronic liver disease patients., Biochem Biophys Rep., 10.1016/j.bbrep.2021.101068, 27, 101068-101068, 2021.07.
2. Tanaka M, Goya T, Suzuki H, Takahashi M, Imoto K, Kurokawa M, Tashiro S, Kuwano A, Okada S, Kato M, Kohjima M, Kotoh K, Ogawa Y., Hypoxic hepatitis with marked elevation of serum ferritin probably due to activation of intrahepatic macrophages: another form of hypoxic hepatitis hitherto not reported?, Acta Gastroenterol Belg., 84, 2, 317-320, 2021.06.
3. Kuwano A, Kurokawa M, Kohjima M, Imoto K, Tashiro S, Suzuki H, Tanaka M, Okada S, Kato M, Ogawa Y., Microcirculatory disturbance in acute liver injury., Exp Ther Med., 21, 6, 596, 2021.06.
4. Suzuki H, Kohjima M, Tanaka M, Goya T, Itoh S, Yoshizumi T, Mori M, Tsuda M, Takahashi M, Kurokawa M, Imoto K,Tashiro S, Kuwano A, Kato M, Okada S, Nakamuta M, Ogawa Y., Metabolic Alteration in Hepatocellular Carcinoma: Mechanism of Lipid Accumulation in Well-Differentiated Hepatocellular Carcinoma., Can J Gastroenterol Hepatol., 2021, 8813410., 2021.02.
5. Okuno H, Ogino H, Ihara E, Nishioka K, Tanaka Y, Chinen T, Kohjima M, Oono T, Tanaka M, Goya T, Fujimori N, Iboshi Y, Gotoda T, Ogawa Y., Discriminant equation using mucosally expressed cytokines and transcription factor for making definite diagnosis of inflammatory bowel disease unclassified., BMC Gastroenterol., 21, 1, 73, 2021.02.
6. Hata K, Suetsugu K, Egashira N, Makihara Y, Itoh S, Yoshizumi T, Tanaka M, Kohjima M, Watanabe H, Masuda S, Ieiri I., Association of lenvatinib plasma concentration with clinical efficacy and adverse events in patients with hepatocellular carcinoma. , Cancer Chemother Pharmacol., 10.1007/s00280-020-04178-x. , 2020.10.
7. Kuwano A, Kohjima M, Suzuki H, Yamasaki A, Ohashi T, Imoto K, Kurokawa M, Morita Y, Kato M, Ogawa Y., Recombinant human soluble thrombomodulin ameliorates acetaminophen-induced liver toxicity in mice., Exp Ther Med., 18, 1323-1330, 2019.07.
8. Ryoko Takei, Tomoaki Inoue, Noriyuki Sonoda, Motoyuki Kohjima, Misato Okamoto, Ryuichi Sakamoto, Toyoshi Inoguchi, Yoshihiro Ogawa, Bilirubin reduces visceral obesity and insulin resistance by suppression of inflammatory cytokines, PloS one, 10.1371/journal.pone.0223302, 14, 10, 2019.01, Objective Although previous studies have reported a negative relationship between serum bilirubin concentration and the development of diabetes mellitus (DM), the relationship between bilirubin and insulin resistance has not been thoroughly assessed. This study was designed to determine the relationships between bilirubin, body fat distribution, and adipose tissue inflammation in patients with type 2 DM and the effect of bilirubin in an obese animal model. Method Body fat distribution was measured using an abdominal dual bioelectrical impedance analyzer in patients with type 2 DM. We also measured glycemic control, lipid profile, serum bilirubin concentration and other clinical characteristics, and determined their relationships with body fat distribution. In the animal study, biliverdin (20 mg/kg daily) was orally administered to high-fat diet (HFD)-induced obese (DIO) mice for 2 weeks, after which intraperitoneal insulin tolerance testing was performed. Then, adipocyte area, adipocytokine expression, and macrophage polarization were evaluated in epididymal adipose tissues. Results In the clinical study, univariate analysis showed that a lower bilirubin concentration was significantly correlated with higher body mass index, waist circumference, triglyceride, uric acid, creatinine, visceral fat area and lower HDL-C. In multivariate analyses, bilirubin concentration significantly correlated with diastolic blood pressure, creatinine, and visceral fat area. However, there was no association between bilirubin concentration and subcutaneous fat area. In the animal study, DIO mice treated with biliverdin had smaller adipocytes than untreated DIO mice and biliverdin improved HFD-induced insulin resistance. Biliverdin treatment reversed the higher gene expression of Cd11c, encoding an M1 macrophage marker, and Tnfa, encoding the proinflammatory cytokine tumor necrosis factor-α, in the adipose tissues of DIO mice. These data suggest biliverdin administration alleviates insulin resistance by ameliorating inflammation and the dysregulation of adipocytokine expression in adipose tissues of DIO mice. Conclusions Bilirubin may protect against insulin resistance by ameliorating visceral obesity and adipose tissue inflammation..
9. Koji Imoto, Motoyuki Kohjima, Tomonobu Hioki, Tomoyuki Kurashige, Miho Kurokawa, Shigeki Tashiro, Hideo Suzuki, Akifumi Kuwano, Masatake Tanaka, Seiji Okada, Masaki Kato, Yoshihiro Ogawa, Clinical Features of Liver Injury Induced by Immune Checkpoint Inhibitors in Japanese Patients, Canadian Journal of Gastroenterology and Hepatology, 10.1155/2019/6391712, 2019, 2019.01, Aim. Immune checkpoint inhibitors (ICIs) have improved the survival rate of patients carrying various malignant neoplasms. Despite their efficacy, ICIs occasionally induce liver injury as an immune-related adverse event (irAE). This study aimed to reveal the clinical features of the hepatic irAE in Japanese patients. Methods. Among 387 patients treated with ICIs, those who developed drug-induced liver injury were investigated. We also describe the histological findings and clinical courses of four patients with hepatic irAE who underwent liver biopsy. Results. Among the 56 patients with all-grade liver injury, only 11 (19.6%) showed hepatocellular-type liver injury, which resembled autoimmune hepatitis. Thirty-four patients (60.7%) developed cholestatic or mixed-type liver injury, although only one patient showed abnormal image findings in the bile duct. Most patients with grade ≤2 liver injury improved spontaneously, while two patients with biliary dysfunction required ursodeoxycholic acid or prednisolone. Among eight patients with grade ≥3 liver injury, three required no immunosuppressants and five were treated with prednisolone (three of five patients required other types of immunosuppressants). Four patients in the case series showed diverse clinical features in terms of hepatotoxic pattern, symptoms, and the interval time between the initiation of immunotherapy and the onset of the hepatic irAE. Conclusions. Our findings suggest that ICIs could cause microscopic biliary disorder without any abnormal image finding. Because the hepatic irAE presents diverse clinical features, liver biopsy is recommended to provide appropriate treatments..
10. Koji Imoto, Motoyuki Kohjima, Tomoyuki Kurashige, Taiji Mutsuki, Shigeki Tashiro, Hideo Suzuki, Akifumi Kuwano, Masaki Kato, Yoshihiro Ogawa, Successful endoscopic treatment of hepatoduodenal fistula formed during sorafenib treatment for hepatocellular carcinoma with duodenal invasion, Acta Hepatologica Japonica, 10.2957/kanzo.60.91, 60, 3, 91-98, 2019.01, Here, we have reported a case of hepatocellular carcinoma (HCC) with a duodenal fistula that was treated with endoscopic procedure in a 79-year-old female patient who was admitted to our hospital with melena. She was diagnosed with recurrent HCC with duodenal bulb invasion and hemorrhage. After 2 months of sorafenib treatment, an improvement in HCC was noted, while a hepatoduodenal fistula was detected at the invasion site. Hepatoduodenal fistula occurring in duodenal invasion of HCC often produces intractable infection and worsens prognosis. We successfully closed the fistula using argon plasma coagulation (APC), following endoscopic injection with fibrin glue and polyglycolic acid sheet coverage. Recent developments in chemotherapy for advanced HCC may provide similar cases with hepatoduodenal fistula occurring in duodenal invasion of HCC and endoscopic treatment could be one of the therapeutic options in such cases..
11. Tomoko Ohashi, Masaki Kato, Akihiro Yamasaki, Akifumi Kuwano, Hideo Suzuki, Motoyuki Kohjima, Yoshihiro Ogawa, Effects of high fructose intake on liver injury progression in high fat diet induced fatty liver disease in ovariectomized female mice, Food and Chemical Toxicology, 10.1016/j.fct.2018.05.006, 118, 190-197, 2018.08, Epidemiology shows that the morbidity of nonalcoholic fatty liver disease (NAFLD) is increased in postmenopausal women and chronic high fructose intake induces NAFLD progression. To analyze the effects of high fructose intake on estrogen deficiency, we evaluated liver disease progression using ovariectomized mice fed with a high fat diet (HFD) for 12 weeks. Hepatic steatosis developed in all HFD groups. Fructose intake significantly increased the liver weight and serum alanine aminotransferase, which was not exacerbated by ovariectomy alone. Ovariectomy enhanced the hepatic inflammatory activity shown by tumor necrosis factor α upregulation in the groups with or without fructose intake. Both fructose intake and ovariectomy increased the hepatocytes with ballooning degeneration and hepatic macrophage infiltration and activated hepatic stellate cells. Coexistence of fructose intake and ovariectomy markedly enhanced liver cell destruction, macrophage accumulation, and progression of fibrosis. Liver damage was ameliorated by 17β-estradiol supplementation. These findings suggest that high fructose intake enhanced the progression of NAFLD in ovariectomized female mice..
12. Takeshi Hirayama, Tadashi Ikegami, Akira Honda, Teruo Miyazaki, Sho Ichiro Yara, Motoyuki Kohjima, Makoto Nakamuta, Yasushi Matsuzaki, Differences in the serum 4β-hydroxycholesterol levels of patients with chronic hepatitis C virus (HCV) infection
A possible impact on the efficacy and safety of interferon (IFN)-free treatment, Internal Medicine, 10.2169/internalmedicine.9479-17, 57, 9, 1219-1227, 2018.01, Objective Since the majority of direct-acting antivirals (DAAs) that are used in the treatment of hepatitis C virus (HCV) infection are mainly metabolized by CYP3A4, it is hypothesized that inter-individual differences in CYP3A4 activity may be associated with the bioavailability of these agents. Methods The level of serum 4β-hydroxycholesterol (4βHC), a surrogate marker of CYP3A4 activity, was determined by LC-MS/MS in samples obtained from patients with HCV infection (CHCs) as well as healthy control subjects (CTLs). Serum samples obtained from patients treated with either asunaprevir/daclatasvir (ASV/DCV) or ombitasvir/paritaprevir/ritonavir (OTV/PTV/r) were used for additional assays. Results The serum 4βHC level in CHCs was significantly higher than that in CTLs, and a gender difference was seen among CHCs. In patients treated with OTV/PTV/r, the serum 4βHC level was observed to gradually decrease during the treatment period. In the cohort treated with ASV/DCV, 4 of 83 patients showed virological treatment failure. In pretreatment testing, an Invader assay detected a low prevalence of resistance-associated variants in these four patients. The average serum concentration of DCV/ASV in the treatment-failed group tended to be lower than that in the sustained virological response (SVR) group. The pretreatment serum 4βHC level in patients with treatment failure was significantly higher than that in patients with an SVR but in whom the prevalence of resistance-associated variants was low in the pretreatment setting. Conclusion The evaluation of CYP3A4 activity by measuring 4βHC before treatment may provide additional information that can potentially be used to select cost- and efficacy-optimized treatment of HCV..
13. Kenichiro Yasutake, Shigemi Koga, Yuka Hokko, Michiko Ikemoto, Yuri Yaguchi, Hironori Sakai, Yusuke Murata, Kenji Ohe, Motoyuki Kohjima, Makoto Nakamuta, Munechika Enjoji, Relevance of the Mini Nutritional Assessment in cirrhotic liver disease patients, Asia Pacific Journal of Clinical Nutrition, 10.6133/apjcn.052017.04, 27, 2, 300-305, 2018.01, Background and Objectives: Malnutrition is an important prognostic factor for patients with liver disease and a novel nutritional assessment tool is required for these patients. The aim of this study was to validate the Mini Nutritional Assessment (MNA) as a nutritional screening tool for patients with liver disease, by comparing MNA scores with other nutrition-related parameters. Methods and Study Design: Patients who were hospitalized at the gastroenterology division of Kyushu and Beppu Medical Center were enrolled. The study included 77 patients with liver disease (male/female, 46/31; mean±SD age, 68.5±10.7 years; liver cirrhosis, 64.9%; liver cancer, 61.0%). Correlations of MNA score at hospital admission with anthropometric parameters and blood test data were evaluated. Results: In patients with liver disease, MNA scores demonstrated that 18 (23.4%) had normal nutritional status, 41 (53.2%) were at risk of malnutrition, and 18 (23.4%) were malnourished, indicating that up to 76.6% of the liver disease group were malnourished. Especially, patients with liver cirrhosis had lower scores of nutritional markers and MNA. The MNA score in liver cirrhotic patients correlated with the following parameters: % arm circumference, % triceps skinfolds, ratio of % maximum grasp strength and arm circumference, maximum grasp strength, arm muscle circumference, calf circumference, serum albumin levels, the controlling nutritional status score, and Onodera's prognostic index, while patients without liver cirrhosis did not show such correlation. Conclusions: MNA scores correlated with nutrition-related data in patients with liver cirrhosis. The MNA is an appropriate tool for nutritional screening assessment in these cirrhotic patients of any etiology..
14. Takeshi Hirayama, Tadashi Ikegami, Akira Honda, Teruo Miyazaki, Sho Ichiro Yara, Motoyuki Kohjima, Makoto Nakamuta, Yasushi Matsuzaki, Differences in the serum 4β-hydroxycholesterol levels of patients with chronic hepatitis C virus (HCV) infection
A possible impact on the efficacy and safety of interferon (IFN)-free treatment, Internal Medicine, 10.2169/internalmedicine.9479-17, 57, 9, 1219-1227, 2018.01, Objective Since the majority of direct-acting antivirals (DAAs) that are used in the treatment of hepatitis C virus (HCV) infection are mainly metabolized by CYP3A4, it is hypothesized that inter-individual differences in CYP3A4 activity may be associated with the bioavailability of these agents. Methods The level of serum 4β-hydroxycholesterol (4βHC), a surrogate marker of CYP3A4 activity, was determined by LC-MS/MS in samples obtained from patients with HCV infection (CHCs) as well as healthy control subjects (CTLs). Serum samples obtained from patients treated with either asunaprevir/daclatasvir (ASV/DCV) or ombitasvir/paritaprevir/ritonavir (OTV/PTV/r) were used for additional assays. Results The serum 4βHC level in CHCs was significantly higher than that in CTLs, and a gender difference was seen among CHCs. In patients treated with OTV/PTV/r, the serum 4βHC level was observed to gradually decrease during the treatment period. In the cohort treated with ASV/DCV, 4 of 83 patients showed virological treatment failure. In pretreatment testing, an Invader assay detected a low prevalence of resistance-associated variants in these four patients. The average serum concentration of DCV/ASV in the treatment-failed group tended to be lower than that in the sustained virological response (SVR) group. The pretreatment serum 4βHC level in patients with treatment failure was significantly higher than that in patients with an SVR but in whom the prevalence of resistance-associated variants was low in the pretreatment setting. Conclusion The evaluation of CYP3A4 activity by measuring 4βHC before treatment may provide additional information that can potentially be used to select cost- and efficacy-optimized treatment of HCV..
15. Kenichiro Yasutake, Shigemi Koga, Yuka Hokko, Michiko Ikemoto, Yuri Yaguchi, Hironori Sakai, Yusuke Murata, Kenji Ohe, Motoyuki Kohjima, Makoto Nakamuta, Munechika Enjoji, Relevance of the Mini Nutritional Assessment in cirrhotic liver disease patients, Asia Pacific Journal of Clinical Nutrition, 10.6133/apjcn.052017.04, 27, 2, 300-305, 2018.01, Background and Objectives: Malnutrition is an important prognostic factor for patients with liver disease and a novel nutritional assessment tool is required for these patients. The aim of this study was to validate the Mini Nutritional Assessment (MNA) as a nutritional screening tool for patients with liver disease, by comparing MNA scores with other nutrition-related parameters. Methods and Study Design: Patients who were hospitalized at the gastroenterology division of Kyushu and Beppu Medical Center were enrolled. The study included 77 patients with liver disease (male/female, 46/31; mean±SD age, 68.5±10.7 years; liver cirrhosis, 64.9%; liver cancer, 61.0%). Correlations of MNA score at hospital admission with anthropometric parameters and blood test data were evaluated. Results: In patients with liver disease, MNA scores demonstrated that 18 (23.4%) had normal nutritional status, 41 (53.2%) were at risk of malnutrition, and 18 (23.4%) were malnourished, indicating that up to 76.6% of the liver disease group were malnourished. Especially, patients with liver cirrhosis had lower scores of nutritional markers and MNA. The MNA score in liver cirrhotic patients correlated with the following parameters: % arm circumference, % triceps skinfolds, ratio of % maximum grasp strength and arm circumference, maximum grasp strength, arm muscle circumference, calf circumference, serum albumin levels, the controlling nutritional status score, and Onodera's prognostic index, while patients without liver cirrhosis did not show such correlation. Conclusions: MNA scores correlated with nutrition-related data in patients with liver cirrhosis. The MNA is an appropriate tool for nutritional screening assessment in these cirrhotic patients of any etiology..
16. Enjoji M, Kohjima M, Ohe K, Yao K., How and why do lipid droplets accumulate in gastric epithelial neoplasm?, Ann Pharmacol Pharmceu, 2017; 2(2):1016-1018. , 2017.09.
17. Masayuki Miyazaki, Masayoshi Yada, Kosuke Tanaka, Takeshi Senjyu, Takeshi Goya, Kenta Motomura, Motoyuki Kohjima, Masaki Kato, Akihide Masumoto, Kazuhiro Kotoh, Efficacy of tolvaptan for the patients with advanced hepatocellular carcinoma, World Journal of Gastroenterology, 10.3748/wjg.v23.i29.5379, 23, 29, 5379-5385, 2017.08, Aim: To investigate the factors influenced the efficacy of tolvaptan (TLV) in liver cirrhosis. Methods: We retrospectively enrolled 61 consecutive patients with refractory hepatic ascites. All of them had been treated with furosemide and spironolactone before admission, and treated with TLV for 7 d in our hospital. The effect of TLV was defined by the rate of body weight loss, and the factors that influenced TLV efficacy were analyzed using multiple regression. Results: Coexistent hepatocellular carcinoma (HCC) was the only significant predictive variable that attenuated the efficacy of TLV. In stratified analysis, high doses of furosemide decreased the efficacy of TLV in patients with HCC, and increased efficacy in those without HCC. In the latter, a high Child-Pugh-Turcotte score had a positive influence and a high concentration of lactate dehydrogenase had a negative influence on the effectiveness of TLV. Conclusion: Development of ascites may differ between patients with liver failure and those with HCC progression. A sufficient preceding dose of furosemide decreases diuretic effect of TLV..
18. Masayuki Miyazaki, Masayoshi Yada, Kosuke Tanaka, Takeshi Senjyu, Takeshi Goya, Kenta Motomura, Motoyuki Kohjima, Masaki Kato, Akihide Masumoto, Kazuhiro Kotoh, Efficacy of tolvaptan for the patients with advanced hepatocellular carcinoma, World Journal of Gastroenterology, 10.3748/wjg.v23.i29.5379, 23, 29, 5379-5385, 2017.08, Aim: To investigate the factors influenced the efficacy of tolvaptan (TLV) in liver cirrhosis. Methods: We retrospectively enrolled 61 consecutive patients with refractory hepatic ascites. All of them had been treated with furosemide and spironolactone before admission, and treated with TLV for 7 d in our hospital. The effect of TLV was defined by the rate of body weight loss, and the factors that influenced TLV efficacy were analyzed using multiple regression. Results: Coexistent hepatocellular carcinoma (HCC) was the only significant predictive variable that attenuated the efficacy of TLV. In stratified analysis, high doses of furosemide decreased the efficacy of TLV in patients with HCC, and increased efficacy in those without HCC. In the latter, a high Child-Pugh-Turcotte score had a positive influence and a high concentration of lactate dehydrogenase had a negative influence on the effectiveness of TLV. Conclusion: Development of ascites may differ between patients with liver failure and those with HCC progression. A sufficient preceding dose of furosemide decreases diuretic effect of TLV..
19. Minae Kawashima, Yuki Hitomi, Yoshihiro Aiba, Nao Nishida, Kaname Kojima, Yosuke Kawai, Hitomi Nakamura, Atsushi Tanaka, Mikio Zeniya, Etsuko Hashimoto, Hiromasa Ohira, Kazuhide Yamamoto, Masanori Abe, Kazuhiko Nakao, Satoshi Yamagiwa, Shuichi Kaneko, Masao Honda, Takeji Umemura, Takafumi Ichida, Masataka Seike, Shotaro Sakisaka, Masaru Harada, Osamu Yokosuka, Yoshiyuki Ueno, Michio Senju, Tatsuo Kanda, Hidetaka Shibata, Takashi Himoto, Kazumoto Murata, Yasuhiro Miyake, Hirotoshi Ebinuma, Makiko Taniai, Satoru Joshita, Toshiki Nikami, Hajime Ota, Hiroshi Kouno, Hirotaka Kouno, Makoto Nakamuta, Nobuyoshi Fukushima, Motoyuki Kohjima, Tatsuji Komatsu, Toshiki Komeda, Yukio Ohara, Toyokichi Muro, Tsutomu Yamashita, Kaname Yoshizawa, Yoko Nakamura, Masaaki Shimada, Noboru Hirashima, Kazuhiro Sugi, Keisuke Ario, Eiichi Takesaki, Atsushi Naganuma, Hiroshi Mano, Haruhiro Yamashita, Kouki Matsushita, Kazuhiko Yamauchi, Fujio Makita, Hideo Nishimura, Kiyoshi Furuta, Naohiro Takahashi, Masahiro Kikuchi, Naohiko Masaki, Tomohiro Tanaka, Sumito Tamura, Akira Mori, Shintaro Yagi, Ken Shirabe, Atsumasa Komori, Kiyoshi Migita, Masahiro Ito, Shinya Nagaoka, Seigo Abiru, Hiroshi Yatsuhashi, Michio Yasunami, Shinji Shimoda, Kenichi Harada, Hiroto Egawa, Yoshihiko Maehara, Shinji Uemoto, Norihiro Kokudo, Hajime Takikawa, Hiromi Ishibashi, Kazuaki Chayama, Masashi Mizokami, Masao Nagasaki, Katsushi Tokunaga, Minoru Nakamura, Genome-wide association studies identify PRKCB as a novel genetic susceptibility locus for primary biliary cholangitis in the Japanese population, Human Molecular Genetics, 10.1093/hmg/ddw406, 26, 3, 650-659, 2017.01, A previous genome-wide association study (GWAS) performed in 963 Japanese individuals (487 primary biliary cholangitis [PBC] cases and 476 healthy controls) identified TNFSF15 (rs4979462) and POU2AF1 (rs4938534) as strong susceptibility loci for PBC. In this study, we performed GWAS in additional 1,923 Japanese individuals (894 PBC cases and 1,029 healthy controls),and combined the results with the previous data. This GWAS, together with a subsequent replication study in an independent set of 7,024 Japanese individuals (512 PBC cases and 6,512 healthy controls), identified PRKCB (rs7404928) as a novel susceptibility locus for PBC (odds ratio [OR] = 1.26, P=4.13×10-9). Furthermore, a primary functional variant of PRKCB (rs35015313) was identified by genotype imputation using a phased panel of 1,070 Japanese individuals from a prospective, general population cohort study and subsequent in vitro functional analyses. These results may lead to improved understanding of the disease pathways involved in PBC, forming a basis for prevention of PBC and development of novel therapeutics..
20. Minae Kawashima, Yuki Hitomi, Yoshihiro Aiba, Nao Nishida, Kaname Kojima, Yosuke Kawai, Hitomi Nakamura, Atsushi Tanaka, Mikio Zeniya, Etsuko Hashimoto, Hiromasa Ohira, Kazuhide Yamamoto, Masanori Abe, Kazuhiko Nakao, Satoshi Yamagiwa, Shuichi Kaneko, Masao Honda, Takeji Umemura, Takafumi Ichida, Masataka Seike, Shotaro Sakisaka, Masaru Harada, Osamu Yokosuka, Yoshiyuki Ueno, Michio Senju, Tatsuo Kanda, Hidetaka Shibata, Takashi Himoto, Kazumoto Murata, Yasuhiro Miyake, Hirotoshi Ebinuma, Makiko Taniai, Satoru Joshita, Toshiki Nikami, Hajime Ota, Hiroshi Kouno, Hirotaka Kouno, Makoto Nakamuta, Nobuyoshi Fukushima, Motoyuki Kohjima, Tatsuji Komatsu, Toshiki Komeda, Yukio Ohara, Toyokichi Muro, Tsutomu Yamashita, Kaname Yoshizawa, Yoko Nakamura, Masaaki Shimada, Noboru Hirashima, Kazuhiro Sugi, Keisuke Ario, Eiichi Takesaki, Atsushi Naganuma, Hiroshi Mano, Haruhiro Yamashita, Kouki Matsushita, Kazuhiko Yamauchi, Fujio Makita, Hideo Nishimura, Kiyoshi Furuta, Naohiro Takahashi, Masahiro Kikuchi, Naohiko Masaki, Tomohiro Tanaka, Sumito Tamura, Akira Mori, Shintaro Yagi, Ken Shirabe, Atsumasa Komori, Kiyoshi Migita, Masahiro Ito, Shinya Nagaoka, Seigo Abiru, Hiroshi Yatsuhashi, Michio Yasunami, Shinji Shimoda, Kenichi Harada, Hiroto Egawa, Yoshihiko Maehara, Shinji Uemoto, Norihiro Kokudo, Hajime Takikawa, Hiromi Ishibashi, Kazuaki Chayama, Masashi Mizokami, Masao Nagasaki, Katsushi Tokunaga, Minoru Nakamura, Genome-wide association studies identify PRKCB as a novel genetic susceptibility locus for primary biliary cholangitis in the Japanese population, Human Molecular Genetics, 10.1093/hmg/ddw406, 26, 3, 650-659, 2017.01, A previous genome-wide association study (GWAS) performed in 963 Japanese individuals (487 primary biliary cholangitis [PBC] cases and 476 healthy controls) identified TNFSF15 (rs4979462) and POU2AF1 (rs4938534) as strong susceptibility loci for PBC. In this study, we performed GWAS in additional 1,923 Japanese individuals (894 PBC cases and 1,029 healthy controls),and combined the results with the previous data. This GWAS, together with a subsequent replication study in an independent set of 7,024 Japanese individuals (512 PBC cases and 6,512 healthy controls), identified PRKCB (rs7404928) as a novel susceptibility locus for PBC (odds ratio [OR] = 1.26, P=4.13×10-9). Furthermore, a primary functional variant of PRKCB (rs35015313) was identified by genotype imputation using a phased panel of 1,070 Japanese individuals from a prospective, general population cohort study and subsequent in vitro functional analyses. These results may lead to improved understanding of the disease pathways involved in PBC, forming a basis for prevention of PBC and development of novel therapeutics..
21. Naoto Kubota, Tetsuya Kubota, Eiji Kajiwara, Tomokatsu Iwamura, Hiroki Kumagai, Taku Watanabe, Mariko Inoue, Iseki Takamoto, Takayoshi Sasako, Katsuyoshi Kumagai, Motoyuki Kohjima, Makoto Nakamuta, Masao Moroi, Kaoru Sugi, Tetsuo Noda, Yasuo Terauchi, Kohjiro Ueki, Takashi Kadowaki, Differential hepatic distribution of insulin receptor substrates causes selective insulin resistance in diabetes and obesity, Nature communications, 10.1038/ncomms12977, 7, 2016.10, Hepatic insulin signalling involves insulin receptor substrates (Irs) 1/2, and is normally associated with the inhibition of gluconeogenesis and activation of lipogenesis. In diabetes and obesity, insulin no longer suppresses hepatic gluconeogenesis, while continuing to activate lipogenesis, a state referred to as 'selective insulin resistance'. Here, we show that 'selective insulin resistance' is caused by the differential expression of Irs1 and Irs2 in different zones of the liver. We demonstrate that hepatic Irs2-knockout mice develop 'selective insulin resistance', whereas mice lacking in Irs1, or both Irs1 and Irs2, develop 'total insulin resistance'. In obese diabetic mice, Irs1/2-mediated insulin signalling is impaired in the periportal zone, which is the primary site of gluconeogenesis, but enhanced in the perivenous zone, which is the primary site of lipogenesis. While hyperinsulinaemia reduces Irs2 expression in both the periportal and perivenous zones, Irs1 expression, which is predominantly in the perivenous zone, remains mostly unaffected. These data suggest that 'selective insulin resistance' is induced by the differential distribution, and alterations of hepatic Irs1 and Irs2 expression..
22. Mari Ohe, Taishi Mutsuki, Takeshi Goya, Shinsaku Yamashita, Takeaki Satoh, Motoyuki Kohjima, Masaki Kato, Portal Vein Thrombosis Repeatedly Observed in a Cirrhotic Patient with Antiphospholipid Antibody Syndrome, Fukuoka igaku zasshi = Hukuoka acta medica, 107, 10, 185-190, 2016.10, Background: Although portal vein thrombosis in cirrhotic patients is frequently observed, the detailed process remains to be clarified, and the role of anticardiolipin antibody in the development of portal vein thrombosis has been controversial.
Case Report: A 52-year-old man, who had been diagnosed with alcoholic cirrhosis of the liver, was admitted to our hospital suffering from dyspnea and ascites. Just after being diagnosed as having antiphospholipid antibody syndrome with lung thrombosis and delivering a positive result for the β 2-glycoprotein I-dependent anticardiolipin antibody, he sustained rupture of the esophageal varices with rapid development of portal vein thrombosis, which resolved under anticoagulant therapy. Two years later, he was admitted again on suspicion of thrombosis because of an elevation in the serum D-dimer level, and computed tomography showed portal and upper mesenteric vein thrombosis. Although immediate anticoagulant therapy resulted in complete recanalization, he suffered the same episode 2 months later, which occurred with re-elevation of the serum D-dimer level.
Conclusion: A positive finding of an anticardiolipin antibody in cirrhotic patients has been considered to be nonspecific and not related to the development of thrombus in the portal vein. This case, however, seems to indicate that cirrhotic patients with the β2-glycoprotein I-dependent anticardiolipin antibody should be regarded as being at high risk for portal vein thrombosis. Monitoring with the serum D-dimer was useful in detecting portal vein thrombosis in its early stage..
23. Naoto Kubota, Tetsuya Kubota, Eiji Kajiwara, Tomokatsu Iwamura, Hiroki Kumagai, Taku Watanabe, Mariko Inoue, Iseki Takamoto, Takayoshi Sasako, Katsuyoshi Kumagai, Motoyuki Kohjima, Makoto Nakamuta, Masao Moroi, Kaoru Sugi, Tetsuo Noda, Yasuo Terauchi, Kohjiro Ueki, Takashi Kadowaki, Differential hepatic distribution of insulin receptor substrates causes selective insulin resistance in diabetes and obesity, Nature communications, 10.1038/ncomms12977, 7, 2016.10, Hepatic insulin signalling involves insulin receptor substrates (Irs) 1/2, and is normally associated with the inhibition of gluconeogenesis and activation of lipogenesis. In diabetes and obesity, insulin no longer suppresses hepatic gluconeogenesis, while continuing to activate lipogenesis, a state referred to as 'selective insulin resistance'. Here, we show that 'selective insulin resistance' is caused by the differential expression of Irs1 and Irs2 in different zones of the liver. We demonstrate that hepatic Irs2-knockout mice develop 'selective insulin resistance', whereas mice lacking in Irs1, or both Irs1 and Irs2, develop 'total insulin resistance'. In obese diabetic mice, Irs1/2-mediated insulin signalling is impaired in the periportal zone, which is the primary site of gluconeogenesis, but enhanced in the perivenous zone, which is the primary site of lipogenesis. While hyperinsulinaemia reduces Irs2 expression in both the periportal and perivenous zones, Irs1 expression, which is predominantly in the perivenous zone, remains mostly unaffected. These data suggest that 'selective insulin resistance' is induced by the differential distribution, and alterations of hepatic Irs1 and Irs2 expression..
24. Mari Ohe, Taishi Mutsuki, Takeshi Goya, Shinsaku Yamashita, Takeaki Satoh, Motoyuki Kohjima, Masaki Kato, Portal Vein Thrombosis Repeatedly Observed in a Cirrhotic Patient with Antiphospholipid Antibody Syndrome, Fukuoka igaku zasshi = Hukuoka acta medica, 107, 10, 185-190, 2016.10, Background: Although portal vein thrombosis in cirrhotic patients is frequently observed, the detailed process remains to be clarified, and the role of anticardiolipin antibody in the development of portal vein thrombosis has been controversial.
Case Report: A 52-year-old man, who had been diagnosed with alcoholic cirrhosis of the liver, was admitted to our hospital suffering from dyspnea and ascites. Just after being diagnosed as having antiphospholipid antibody syndrome with lung thrombosis and delivering a positive result for the β 2-glycoprotein I-dependent anticardiolipin antibody, he sustained rupture of the esophageal varices with rapid development of portal vein thrombosis, which resolved under anticoagulant therapy. Two years later, he was admitted again on suspicion of thrombosis because of an elevation in the serum D-dimer level, and computed tomography showed portal and upper mesenteric vein thrombosis. Although immediate anticoagulant therapy resulted in complete recanalization, he suffered the same episode 2 months later, which occurred with re-elevation of the serum D-dimer level.
Conclusion: A positive finding of an anticardiolipin antibody in cirrhotic patients has been considered to be nonspecific and not related to the development of thrombus in the portal vein. This case, however, seems to indicate that cirrhotic patients with the β2-glycoprotein I-dependent anticardiolipin antibody should be regarded as being at high risk for portal vein thrombosis. Monitoring with the serum D-dimer was useful in detecting portal vein thrombosis in its early stage..
25. Masayoshi Yada, Masayuki Miyazaki, Kenta Motomura, Akihide Masumoto, Makoto Nakamuta, Motoyuki Kohjima, Rie Sugimoto, Yoshifusa Aratake, Nobuhiko Higashi, Shusuke Morizono, Shinichiro Takao, Naoki Yamashita, Takeaki Satoh, Shinsaku Yamashita, Masami Kuniyoshi, Kazuhiro Kotoh, The prognostic role of lactate dehydrogenase serum levels in patients with hepatocellular carcinoma who are treated with sorafenib
The influence of liver fibrosis, Journal of Gastrointestinal Oncology, 10.21037/jgo.2016.03.10, 7, 4, 615-623, 2016.08, Background: Serum lactate dehydrogenase (LDH) levels could be a prognostic factor for sorafenib-treated patients with several types of solid tumor because it reflects hypoxic circumstances in aggressive tumors. For hepatocellular carcinoma (HCC), however, the prognostic role of LDH has been controversial. Liver fibrosis can potentially cause hypoxia in the liver, which has not been previously studied in the patients with advanced HCC. Thus, we aimed to analyze the prognostic role of LDH based on the degree of fibrosis. Methods: Eighty-nine consecutive patients with HCC (Child-Pugh class A) who were treated using sorafenib were enrolled into this study. Pretreatment characteristics and changes in hepatic functional tests based on early response to sorafenib and serum LDH levels were analyzed. The degree of fibrosis was estimated using the aspartate aminotransferase (AST) to platelet ratio index (APRI), and the tumor response was evaluated after 3 months of sorafenib treatment. Results: Overall, five patients discontinued sorafenib within 4 weeks. For the other 84 patients, those with progressive disease (PD) had significantly high pretreatment LDH levels, which correlated with the APRI score but not with the tumor stage. Multivariate logistic analysis revealed that older age and lower pretreatment LDH levels were independent prognostic factors for a better response to sorafenib. In patients who discontinued sorafenib early, three experienced acute liver failure accompanied with an increase in serum LDH. Conclusions: We demonstrated that baseline serum LDH levels in HCC patients were affected by liver fibrosis but not by the tumor stage, and these LDH levels could be a marker for early response to sorafenib. A marked increase in serum LDH levels during sorafenib administration might also indicate subsequent acute liver failure. Close observation of serum LDH levels before and during sorafenib treatment could be useful in managing treatment of patients receiving this therapy..
26. Motoyuki Kohjima, Miho Kurokawa, Munechika Enjoji, Tsuyoshi Yoshimoto, Tsukasa Nakamura, Tomoko Ohashi, Kunitaka Fukuizumi, Naohiko Harada, Yusuke Murata, Kazuhisa Matsunaga, Masaki Kato, Kazuhiro Kotoh, Makoto Nakamuta, Analysis of renal function during telaprevir-based triple therapy for chronic hepatitis C, Experimental and Therapeutic Medicine, 10.3892/etm.2016.3133, 11, 5, 1781-1787, 2016.05, Telaprevir (TVR) is used for the treatment of chronic hepatitis C in a combination therapy with pegylated-interferon and ribavirin. Although renal dysfunction is one of the critical adverse outcomes of this treatment, little is known regarding the mechanism of its onset. The present study assessed the association of renal function with TVR dose and viral response. Hematological, biochemical, urinary and virological parameters of renal function were examined during the TVR-based triple therapy of patients infected with hepatitis C virus (HCV) genotype 1b. Serum creatinine levels were increased and the estimated glomerular filtration rate (eGFR) was decreased in every patient during TVR administration, but these values recovered to normal levels following cessation of TVR. Fractional excretion of sodium was <1% at days 3 and 7, appearing similar regardless of baseline renal function. Urinary β2-microglobulin levels were elevated and were significantly higher in patients with renal dysfunction, as compared with those not exhibiting renal dysfunction (P<0.05). The reduction in renal function was milder in patients treated with a reduced TVR dose, and these patients had a significantly lower risk of developing renal dysfunction (P<0.05). Using a multivariate analysis, TVR dose and eGFR at the initiation of treatment were identified as significant contributory factors in the development of renal dysfunction. Reduction in TVR dose did not lead to a significant increase in the viral kinetics of HCV or detrimental effects on the sustained viral response (SVR) rate. It is hypothesized that renal dysfunction during TVR treatment is caused by damage of the renal tubule, in addition to pre-renal dysfunction, and that reduction in TVR dose reduces the rate of renal dysfunction without causing a significant decrease in the SVR rate..
27. Munechika Enjoji, Motoyuki Kohjima, Kensei Ohtsu, Kazuhisa Matsunaga, Yusuke Murata, Makoto Nakamuta, Kentaro Imamura, Hiroshi Tanabe, Akinori Iwashita, Takashi Nagahama, Kenshi Yao, Intracellular mechanisms underlying lipid accumulation (white opaque substance) in gastric epithelial neoplasms
A pilot study of expression profiles of lipid-metabolism-associated genes, Journal of Gastroenterology and Hepatology (Australia), 10.1111/jgh.13216, 31, 4, 776-781, 2016.04, Background and Aim:: White opaque substance (WOS) is a novel endoscopic finding in gastric neoplasms, indicating the intracellular accumulation of lipid droplets (LDs). However, gastric lipid metabolism has not been extensively investigated, even in normal mucosa. We investigated the expression profiles of lipid-metabolism-associated genes in gastric neoplasms. Methods:: Thirty-four patients with early gastric cancer or adenoma were enrolled in this study. Paired biopsy samples from tumor and adjacent non-tumor areas were obtained and analyzed by real-time polymerase chain reaction. Endoscopically resected specimens were evaluated histopathologically. Results:: Genes associated with β-oxidation (peroxisome proliferator-activated receptor α, carnitine palmitoyltransferase 1A, and hydroxyacyl-CoA dehydrogenase), lipoprotein excretion (apolipoprotein B, microsomal triglyceride transfer protein, and acyl-CoA:cholesterol acyltransferase 2), fatty acid transport (fatty acid-binding protein), construction of triglycerides in the endoplasmic reticulum (acyl-CoA:diacylglycerol acyltransferase 1), and LD degradation/lipolysis (comparative gene identification-58, adipose triglyceride lipase) were significantly downregulated in neoplasms compared with non-tumor areas. Pyruvate dehydrogenase lipoamide kinase isozyme 4 (negative regulator of glycolysis) and adipophilin (LD surface component) were also repressed. Conversely, expression levels of genes associated with de novo lipogenesis (sterol regulatory element-binding protein 1c, acyl-CoA:diacylglycerol acyltransferase 2) were significantly enhanced in neoplasms. There was no significant difference in gene expression levels between carcinomas and adenomas, or between WOS-positive and WOS-negative neoplasms. Conclusion:: Gene expression profiles in neoplasms suggest a predominance of lipid storage (lipogenesis/LD formation) over consumption (β-oxidation/excretion/lipolysis). Lipid accumulation and WOS in gastric epithelial neoplasms may be caused by impaired mitochondrial oxidation, lipoprotein excretion, and LD degradation..
28. Motoyuki Kohjima, Kenta Motomura, Toshimasa Koyanagi, Seiya Tada, Naoki Yamashita, Takeaki Sato, Syoji Nagase, Masaki Kato, Kazuhiro Kotoh, Makoto Nakamuta, Therapeutic safety and effectiveness of dual oral therapy with daclatasvir and asunaprevir for chronic HCV patients with renal dysfunction, Acta Hepatologica Japonica, 10.2957/kanzo.57.142, 57, 3, 142-145, 2016.04, We investigated the safety and effectiveness of the dual oral therapy with Daclatasvir (DCV) and Asunaprevir (ASV) for patients with renal dysfunctioa This study enrolled 682 HCV genotype lb patients treated with DCV and ASV and included 83 patients with eGFR less than 50 m//min/1.73 m2, and 17 patients with eGFR less than 30 m//min/L73 m2. Serum creatinine and eGFR were not altered during the treatment and the rate of complications and viral response was similar between patients with renal dysfunction and patients without renal dysfunction. The treatment with DCV and ASV was well tolerated and safety enough even for the HCV patients with renal dysfunction and could be productive therapeutic option for aged patients that accounts many of patients with HCV genotype lb infection in Japan..
29. Miyuki Kuwayama, Keita Uchino, Kotoe Takayoshi, Masato Komoda, Motoyuki Kohjima, Makoto Nakamuta, Seiya Momosaki, Hitoshi Kusaba, Koichi Akashi, Eishi Baba, Immunosuppressant therapy successfully improved regorafenib-induced severe hepatic injury in a patient with metastatic gastrointestinal stromal tumor
A case report, Oncology Letters, 10.3892/ol.2015.3853, 11, 1, 85-88, 2016.01, A 75-year-old man diagnosed with ileal gastrointestinal tumor with peritoneal dissemination was subjected to salvage treatment with regorafenib at 120 mg/day. Following the initiation of the treatment, liver dysfunction appeared on day 28, and continued to worsen despite termination of the treatment. Since no increase in the levels of serum immunoglobulins of the patient was observed, and negative results were obtained for the analysis of viral markers and autoantibodies, a diagnosis of regorafenib-induced hepatitis was suggested. In consequence, the patient received steroid pulse therapy and continuous administration of prednisolone, without sufficient improvement. Liver biopsy revealed interface hepatitis with prominent plasma cell infiltration, suggesting regorafenib-induced autoimmune hepatitis. The patient was then administered azathioprine and prednisolone, which improved the hepatic injury. The present case represents the first report of successful treatment of regorafenib-induced severe hepatic injury by the use of an immunosuppressant..
30. Munechika Enjoji, Motoyuki Kohjima, Makoto Nakamuta, Lipid metabolism and the liver, The Liver in Systemic Diseases, 10.1007/978-4-431-55790-6_6, 105-122, 2016.01, Generally, lipid metabolism in liver hepatocytes can be summarized by three processes: (1) acquisition of lipids, including uptake of lipids and fatty acids, and fatty acid synthesis (de novo lipogenesis); (2) lipid storage, including triglyceride synthesis and the formation of lipid droplets; and (3) lipid consumption, including fatty acid degradation (lipolysis), β-oxidation, and the secretion of very low-density lipoproteins. Many liver diseases are accompanied by disorders in lipid metabolism, with the latter often involving the clinical condition of these patients. This chapter summarizes current understanding of hepatic lipid metabolism, including cholesterol and phospholipid metabolism, and associated factors. In addition, we make reference to metabolic disturbance in some liver diseases..
31. Makoto Nakamuta, Motoyuki Kohjima, Effects of telaprevir (TVR) on cholesterol metabolism, Nihon rinsho. Japanese journal of clinical medicine, 73, 199-203, 2015.12.
32. Ryu Takeya, Noriko Ueno, Keiichiro Kami, Masahiko Taura, Motoyuki Kohjima, Tomoko Izaki, Hiroyuki Nunoi, Hideki Sumimoto, Additions and corrections
Novel human homologues of p47phoxand p67phox participate in activation of superoxide-producing NADPH oxidases (Journal of Biological Chemistry (2003) 278, (25234-25246)), Journal of Biological Chemistry, 10.1074/jbc.A114.212856, 290, 10, 2015.03.
33. Motoyuki Kohjima, Tsuyoshi Yoshimoto, Munechika Enjoji, Nobuyoshi Fukushima, Kunitaka Fukuizumi, Tsukasa Nakamura, Miho Kurokawa, Nao Fujimori, Yusuke Sasaki, Yasushi Shimonaka, Yusuke Murata, Susumu Koyama, Ken Kawabe, Kazuhiro Haraguchi, Yorinobu Sumida, Naohiko Harada, Masaki Kato, Kazuhiro Kotoh, Makoto Nakamuta, Hepcidin/ferroportin expression levels involve efficacy of pegylated-interferon plus ribavirin in hepatitis C virus-infected liver, World Journal of Gastroenterology, 10.3748/wjg.v21.i11.3291, 21, 11, 3291-3299, 2015.03, AIM: To investigate the relationship between the ironmetabolismrelated gene expression profiles and efficacy of antiviral therapy in chronic hepatitis C patients. METHODS: The hepatic expression profile of ironmetabolismrelated genes was analyzed and its association with virological response to pegylated-interferon plus ribavirin combination therapy was evaluated. A hundred patients with chronic hepatitis C (genotype1b, n = 50; genotype 2, n = 50) were enrolled and retrospectively analyzed. Liver biopsy samples were subjected to quantitative polymerase chain reaction for iron-metabolismrelated genes and protein expression (Western blotting analysis) for ferroportin. As a control, normal liver tissue was obtained from 18 living donors of liver transplantation. Serum hepcidin level was measured by sensitive liquid chromatography/electrospray ionization tandem mass spectrometry. RESULTS: Iron overload is associated with liver damage by increasing oxidative stress and hepatitis C virus (HCV) is reported to induce iron accumulation in hepatocytes in vivo. Conversely, iron administration suppresses HCV replication in vitro. Therefore, the association between HCV infection and iron metabolism remains unclear. Compared with controls, patients had significantly higher gene expression for transferrin, iron-regulatory proteins 1 and 2, divalent metal transporter 1, and ferroportin, but similar for transferrin receptors 1 and 2, and hepcidin. When the expression profiles were compared between sustained virological response (SVR) and non-SVR patients, the former showed significantly lower transcription and protein expression of hepcidin and ferroportin. Expression of hepcidin-regulating genes, BMPR1, BMPR2, and hemojuvelin, was significantly increased, whereas BMP2 was decreased in HCV-infected liver. BMPR2 and hemojuvelin expression was significantly lower in the SVR than non-SVR group. HCV infection affects the expression of iron-metabolism-related genes, leading to iron accumulation in hepatocytes. CONCLUSION: Decreased expression of hepcidin and ferroportin in SVR patients indicates the importance of hepatocytic iron retention for viral response during pegylated-interferon plus ribavirin treatment..
34. Motoyuki Kohjima, Munechika Enjoji, Ryoko Yada, Tsuyoshi Yoshimoto, Tsukasa Nakamura, Kunitaka Fukuizumi, Nobuyoshi Fukushima, Yusuke Murata, Manabu Nakashima, Masaki Kato, Kazuhiro Kotoh, Ken Shirabe, Yoshihiko Maehara, Atsushi Nakajima, Yuichi Nozaki, Akira Honda, Yasushi Matsuzaki, Makoto Nakamuta, Pathophysiological analysis of primary biliary cirrhosis focusing on choline/phospholipid metabolism, Liver International, 10.1111/liv.12526, 35, 3, 1095-1102, 2015.03, Background and Aims: Injury to biliary epithelial cells caused by disorders in bile composition may be the initial step in the pathogenesis of primary biliary cirrhosis (PBC). We therefore examined choline/phospholipid metabolism in livers of patients with PBC. Methods: Hepatic levels of mRNA encoded by choline metabolism-related genes in early stage PBC patients were quantified by real-time RT-PCR. Serum cholesterol and triglyceride concentrations in each lipoprotein compartment and serum/tissue choline levels were also measured. OCT1 expression was quantified by genotype (rs683369 and rs622342). Results: Serum choline concentrations were significantly higher in PBC patients than in normal individuals, with the concentrations in the former lowered by treatment with fibrates. Hepatic choline levels were markedly lower in PBC patients than in controls. The levels of expression of genes associated with choline uptake (OCT1 and CTL1), phosphatidylcholine synthesis (PEMT and BHMT), and phosphatidylcholine transport (MDR3) were significantly upregulated in PBC compared with control livers. Serum cholesterol concentrations and the cholesterol/triglyceride ratio in serum very low density lipoprotein were markedly higher in PBC patients than in controls. In PBC liver, OCT1 protein levels were lower in patients with minor (CG/GG at rs683369 and/or CC at rs622342) than major (CC at rs683369 and AA at rs622342) genotypes of the OCT1 gene. Conclusion: During early stage PBC, hepatocellular choline uptake and PC synthesis become dysregulated. OCT1 genotypes may influence the pathogenesis of PBC..
35. Takumi Kawaguchi, Motoyuki Kohjima, Tatsuki Ichikawa, Masataka Seike, Yasushi Ide, Toshihiko Mizuta, Koichi Honda, Kazuhiko Nakao, Makoto Nakamuta, Michio Sata, The morbidity and associated risk factors of cancer in chronic liver disease patients with diabetes mellitus
a multicenter field survey, Journal of Gastroenterology, 10.1007/s00535-014-0968-5, 50, 3, 333-341, 2015.01, Background and aims: Diabetes mellitus is associated with various cancers; however, little is known of the relationship between cancer and diabetes in chronic liver disease (CLD) patients. The aim of this study is to investigate the morbidity and associated factors of cancer, including the use of anti-diabetics, in CLD patients with diabetes.
Patients and methods: We performed a multicenter survey in 2012 and 478 CLD patients with diabetes were enrolled (age 64.3 ± 12.1 years, female/male 187/291). A frequency analysis of cancer and antidiabetic use was performed. Independent factors for cancer were analyzed using logistic regression and decision-tree analysis.
Results: The morbidity of cancer was 33.3 %. Hepatocellular carcinoma (HCC) and extra-hepatic cancer were diagnosed in 24.7 and 11.3 % of enrolled patients, respectively. The frequency of antidiabetic use was 66.5 %. Of prescribed antidiabetics, 39 % were dipeptidyl-peptidase 4 inhibitors; however, their use was not significantly associated with cancer. In contrast, the use of exogenous insulin (OR 2.21; 95 % CI 1.16–4.21, P = 0.0165) and sulfonylurea (OR 2.08; 95 % CI 1.05–3.97, P = 0.0353) were independently associated with HCC and extra-hepatic cancer, respectively. In decision-tree analysis, exogenous insulin and sulfonylurea were also identified as a divergence factor for HCC and extra-hepatic cancer, respectively.
Conclusions: We found a high morbidity of not only HCC, but also extra-hepatic cancer in CLD patients with diabetes. We also showed a possible association between the use of antidiabetics and the morbidity of cancer. Thus, a large-scale cohort study is needed to establish a therapeutic strategy for diabetes to suppress carcinogenesis in CLD patients..
36. Minako Imamura, Benny Hung Junn Chang, Motoyuki Kohjima, Ming Li, Byounghoon Hwang, Heinrich Taegtmeyer, Robert A. Harris, Lawrence Chan, MondoA deficiency enhances sprint performance in mice, Biochemical Journal, 10.1042/BJ20140530, 464, 35-48, 2014.11, MondoA is a basic helix-loop-helix (bHLH)/leucine zipper (ZIP) transcription factor that is expressed predominantly in skeletal muscle. Studies in vitro suggest that the Max-like protein X (MondoA:Mlx) heterodimer senses the intracellular energy status and directly targets the promoter region of thioredoxin interacting protein (Txnip) and possibly glycolytic enzymes. We generated MondoA-inactivated (MondoA-/-) mice by gene targeting. MondoA-/- mice had normal body weight at birth, exhibited normal growth and appeared to be healthy. However, they exhibited unique metabolic characteristics. MondoA-/- mice built up serum lactate and alanine levels and utilized fatty acids for fuel during exercise. Gene expression and promoter analysis suggested that MondoA functionally represses peroxisomeproliferatoractivated receptor γ co-activator-1α (PGC-1α)-mediated activation of pyruvate dehydrogenase kinase 4 (PDK-4) transcription. PDK4 normally down-regulates the activity of pyruvate dehydrogenase, an enzyme complex that catalyses the decarboxylation of pyruvate to acetyl-CoA for entry into the Krebs cycle; in the absence of MondoA, pyruvate is diverted towards lactate and alanine, both products of glycolysis. Dynamic testing revealed that MondoA-/- mice excel in sprinting as their skeletal muscles display an enhanced glycolytic capacity. Our studies uncover a hitherto unappreciated function of MondoA in fuel selection in vivo. Lack of MondoA results in enhanced exercise capacity with sprinting..
37. Tadashi Ikegami, Akira Honda, Teruo Miyazaki, Motoyuki Kohjima, Makoto Nakamuta, Yasushi Matsuzaki, Increased serum oxysterol concentrations in patients with chronic hepatitis C virus infection, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2014.01.176, 446, 3, 736-740, 2014.04, Oxidative stress and dysregulated cholesterol metabolism are characteristic features of chronic hepatitis C virus infection (CHC). Therefore, we analyzed serum oxysterol profiles in CHC patients and examined the significance of oxysterols in CHC. The concentrations of 7α-hydroxycholesterol, 4β-hydroxycholesterol and 25-hydroxycholesterol as determined by LC-ESI-MS/MS were significantly elevated by +236%, +29% and +44%, respectively, in CHC patients compared with controls. Moreover, the elevated levels were significantly decreased by anti-viral therapy using PEGylated-interferon and ribavirin for 3 months. In contrast, 24S-hydroxycholesterol, 27-hydroxycholesterol and 7α-hydroxy-4-cholesten-3-one concentrations were not affected by CHC or anti-viral treatment. These results suggest that some oxysterols that are elevated in CHC are produced by cholesterol autoxidation due to oxidative stress or inflammation in the liver. Oxysterols may represent novel targets for the inhibition of disease progression and the prevention of hepatocarcinogenesis in CHC patients..
38. Kenichiro Yasutake, Motoyuki Kohjima, Kazuhiro Kotoh, Manabu Nakashima, Makoto Nakamuta, Munechika Enjoji, Dietary habits and behaviors associated with nonalcoholic fatty liver disease, World Journal of Gastroenterology, 10.3748/wjg.v20.i7.1756, 20, 7, 1756-1767, 2014.02, Nonalcoholic fatty liver disease (NAFLD) is one of the most frequent causes of health problems in Western (industrialized) countries. Moreover, the incidence of infantile NAFLD is increasing, with some of these patients progressing to nonalcoholic steatohepatitis. These trends depend on dietary habits and life-style. In particular, overeating and its associated obesity affect the development of NAFLD. Nutritional problems in patients with NAFLD include excess intake of energy, carbohydrates, and lipids, and shortages of polyunsaturated fatty acids, vitamins, and minerals. Although nutritional therapeutic approaches are required for prophylaxis and treatment of NAFLD, continuous nutrition nutrition therapy is difficult for many patients because of their dietary habits and lifestyle, and because the motivation for treatment differs among patients. Thus, it is necessary to assess the nutritional background and to identify nutritional problems in each patient with NAFLD. When assessing dietary habits, it is important to individually evaluate those that are consumed excessively or insufficiently, as well as inappropriate eating behaviors. Successful nutrition therapy requires patient education, based on assessments of individual nutrients, and continuing the treatment. In this article, we update knowledge about NAFLD, review the important aspects of nutritional assessment targeting treatment success, and present some concrete nutritional care plans which can be applied generally..
39. Yuki Ohishi, Makoto Nakamuta, Naoko Ishikawa, Ohki Saitoh, Hitomi Nakamura, Yoshihiro Aiba, Atsumasa Komori, Kiyoshi Migita, Hiroshi Yatsuhashi, Nobuyoshi Fukushima, Motoyuki Kohjima, Tsuyoshi Yoshimoto, Kunitaka Fukuizumi, Makoto Ishibashi, Takashi Nishino, Ken Shirabe, Akinobu Taketomi, Yoshihiko Maehara, Hiromi Ishibashi, Minoru Nakamura, Genetic polymorphisms of OCT-1 confer susceptibility to severe progression of primary biliary cirrhosis in Japanese patients, Journal of gastroenterology, 10.1007/s00535-013-0795-0, 49, 2, 332-342, 2014.02, Background: To identify the genetic factors involved in the pathogenesis of primary biliary cirrhosis (PBC), we focused on the organic cation transporter 1 (OCT1/SLC22A1), which is closely associated with phosphatidylcholine synthesis in hepatocytes. Methods: We selected four (rs683369, rs2282143, rs622342 and rs1443844) OCT-1 single nucleotide polymorphisms (SNPs), and genotyped these SNPs using the TaqMan probe method in 275 Japanese PBC patients and 194 gender-matched, healthy volunteers as controls. Results: The Chi-square test revealed that the rs683369 variant allele (G) was associated with insusceptibility to PBC development [P = 0.009, odds ratio (OR) 0.60, 95 % confidence interval (CI) 0.40-0.88] in an allele model, and that the rs683369 variant allele (G) was associated with jaundice-type progression in a minor allele dominant genotype model (P = 0.032, OR 3.10, 95 % CI 1.05-9.14). The OCT-1 rs2282143 variant (T) and rs622342 variant (C) were also associated with jaundice-type progression in a minor allele recessive genotype model (P = 0.0002, OR 10.58, 95 % CI 2.36-47.54, and P = 0.006, OR 7.84, 95 % CI 1.39-44.36, respectively). Furthermore, the association of OCT-1 rs683369 and rs622342 with susceptibility to jaundice-type progression was confirmed by a replication study with a distinct set of PBC patients who underwent liver transplantation. Conclusions: The present study is the first report on the association of OCT-1 genetic polymorphisms with the overall development and jaundice-type progression of PBC..
40. Kenichiro Yasutake, Motoyuki Kohjima, Makoto Nakamuta, Kazuhiro Kotoh, Yusuke Murata, Munechika Enjoji, [Probiotic nutrition therapy for nonalcoholic fatty liver disease]., Fukuoka igaku zasshi = Hukuoka acta medica, 105, 2, 42-47, 2014.01.
41. Motoyuki Kohjima, Tsung Huang Tsai, Bryan C. Tackett, Sundararajah Thevananther, Lan Li, Benny Hung Junn Chang, Lawrence Chan, Delayed liver regeneration after partial hepatectomy in adipose differentiation related protein-null mice, Journal of Hepatology, 10.1016/j.jhep.2013.07.025, 59, 6, 1246-1254, 2013.12, Background & Aims Adult hepatocytes undergo cell cycle progression and proliferation in response to partial hepatectomy (PH). Transient lipid accumulation within hepatocytes preceding the peak proliferative phase is a characteristic feature of regenerating livers. However, the molecular mediators and mechanisms responsible for lipid accumulation in regenerating livers are not well understood. Adipose differentiation related protein (ADRP; Plin2) regulates hepatic triglyceride storage and Plin2-deficient (Plin2-/-) mice have significantly reduced triglyceride (TG) content in the liver. We sought to determine the functional significance of PLIN2 in liver regeneration in response to PH and toxic liver injury and examined whether absence of Plin2 expression modulates hepatocyte proliferation and liver regeneration. Methods We subjected wild-type (WT) and Plin2-/- mice to 70% PH or acute carbon tetrachloride (CCL4) treatment and examined the hepatic lipid content, the expression profile of lipid metabolism-related genes, the rate of cellular proliferation and the dynamics of liver regeneration in the treated animals. Results In response to PH, Plin2-/- mice showed decreased hepatic triglyceride accumulation and delayed cell cycle progression, which was associated with impaired liver regeneration. Fatty acid (FA) synthesis and lipid transfer gene expression profile were comparable between Plin2-/- and wild-type mice, while VLDL secretion rate was higher in the Plin2 -/- mice. Downregulated β-oxidation and reduced cytosolic FA level in Plin2-/- mice may have contributed to the attenuation of the liver regeneration capacity in these animals. In parallel experiments, we also observed attenuated hepatic lipid accumulation and proliferation in response to CCl4-mediated acute toxic liver injury in Plin2-/- mice. Conclusions We conclude that PLIN2-mediated lipid accumulation and utilization by the liver is important for efficient liver regeneration in response to PH and toxic liver injury..
42. Kiyoshi Migita, Minoru Nakamura, Seigo Abiru, Yuka Jiuchi, Shinya Nagaoka, Atsumasa Komori, Satoru Hashimoto, Shigemune Bekki, Kazumi Yamasaki, Tatsuji Komatsu, Masaaki Shimada, Hiroshi Kouno, Taizo Hijioka, Motoyuki Kohjima, Makoto Nakamuta, Michio Kato, Kaname Yoshizawa, Hajime Ohta, Yoko Nakamura, Eiichi Takezaki, Hideo Nishimura, Takeaki Sato, Keisuke Ario, Noboru Hirashima, Yukio Oohara, Atsushi Naganuma, Toyokichi Muro, Hironori Sakai, Eiji Mita, Kazuhiro Sugi, Haruhiro Yamashita, Fujio Makita, Hiroshi Yatsuhashi, Hiromi Ishibashi, Michio Yasunami, Association of STAT4 Polymorphisms with Susceptibility to Type-1 Autoimmune Hepatitis in the Japanese Population, PloS one, 10.1371/journal.pone.0071382, 8, 8, 2013.08, Background/Aims:Recent studies demonstrated an association of STAT4 polymorphisms with autoimmune diseases including systemic lupus erythematosus and rheumatoid arthritis, indicating multiple autoimmune diseases share common susceptibility genes. We therefore investigated the influence of STAT4 polymorphisms on the susceptibility and phenotype of type-1 autoimmune hepatitis in a Japanese National Hospital Organization (NHO) AIH multicenter cohort study.Methodology/Principal Findings:Genomic DNA from 460 individuals of Japanese origin including 230 patients with type-1 autoimmune hepatitis and 230 healthy controls was analyzed for two single nucleotide polymorphisms in the STAT4 gene (rs7574865, rs7582694). The STAT4 rs7574865T allele conferred risk for type-1 autoimmune hepatitis (OR = 1.61, 95% CI = 1.23-2.11; P = 0.001), and patients without accompanying autoimmune diseases exhibited an association with the rs7574865T allele (OR = 1.50, 95%CI = 1.13-1.99; P = 0.005). Detailed genotype-phenotype analysis of type-1 autoimmune hepatitis patients with (n = 44) or without liver cirrhosis (n = 186) demonstrated that rs7574865 was not associated with the development of liver cirrhosis and phenotype (biochemical data and the presence of auto-antibodies).Conclusions/Significance:This is the first study to show a positive association between a STAT4 polymorphism and type-1 autoimmune hepatitis, suggesting that autoimmune hepatitis shares a gene commonly associated with risk for other autoimmune diseases..
43. Motoyuki Kohjima, Munechika Enjoji, Tsuyoshi Yoshimoto, Ryoko Yada, Tatsuya Fujino, Yoko Aoyagi, Nobuyoshi Fukushima, Kunitaka Fukuizumi, Naohiko Harada, Masayoshi Yada, Masaki Kato, Kazuhiro Kotoh, Manabu Nakashima, Naoya Sakamoto, Yasuhito Tanaka, Makoto Nakamuta, Add-on therapy of pitavastatin and eicosapentaenoic acid improves outcome of peginterferon plus ribavirin treatment for chronic hepatitis C, Journal of Medical Virology, 10.1002/jmv.23464, 85, 2, 250-260, 2013.02, Despite the use of pegylated-interferon (peg-IFN) plus ribavirin combination therapy, many patients infected with hepatitis C virus (HCV)-1b remain HCV-positive. To determine whether addition of pitavastatin and eicosapentaenoic acid (EPA) is beneficial, the "add-on" therapy option (add-on group) was compared retrospectively with unmodified peg-IFN/ribavirin therapy (standard group). Association of host- or virus-related factors with sustained virological response was assessed. In HCV replicon cells, the effects of pitavastatin and/or EPA on HCV replication and expression of innate-immunity- and lipid-metabolism-associated genes were investigated. In patients infected with HCV-1b, sustained virological response rates were significantly higher in the add-on than standard group. In both groups, sustained virological response rates were significantly higher in patients with genotype TT of IL-28B (rs8099917) than in those with non-TT genotype. Among the patients with non-TT genotype, sustained virological response rates were markedly higher in the add-on than standard group. By multivariate analysis, genome variation of IL28B but not add-on therapy remained as a predictive factor of sustained virological response. In replicon cells, pitavastatin and EPA suppressed HCV replication. Activation of innate immunity was obvious in pitavastatin-treated cells and EPA suppressed the expression of sterol regulatory element binding protein-1c and low-density lipoprotein receptor. Addition of pitavastatin and EPA to peg-IFN/ribavirin treatment improved sustained virological response in patients infected with HCV-1b. Genotype variation of IL-28B is a strong predictive factor in add-on therapy. J. Med. Virol. 85:250-260, 2013. © 2012 Wiley Periodicals, Inc..
44. Munechika Enjoji, Kenichiro Yasutake, Motoyuki Kohjima, Makoto Nakamuta, Nutrition and alcoholic and nonalcoholic fatty liver disease
The significance of cholesterol, Alcohol, Nutrition, and Health Consequences, 10.1007/978-1-62703-047-2_40, 523-532, 2013.01.
45. Kenichiro Yasutake, Motoyuki Kohjima, Manabu Nakashima, Kazuhiro Kotoh, Makoto Nakamuta, Munechika Enjoji, Nutrition therapy for liver diseases based on the status of nutritional intake, Clinical Nutrition The Interface Between Metabolism, Diet, and Disease, 10.1201/b16308, 55-74, 2013.01, The liver is one of the main organs of nutritional metabolism, including protein synthesis, glycogen storage, and detoxification. These functions become damaged to a greater or lesser extent in patients with liver diseases, resulting in various metabolic disorders, and their disturbed nutritional condition is associated with disease progression. Therefore, dietary counseling and nutritional intervention can support other medical treatments in some liver diseases..
46. Kenichiro Yasutake, Motoyuki Kohjima, Manabu Nakashima, Kazuhiro Kotoh, Makoto Nakamuta, Munechika Enjoji, Nutrition therapy for liver diseases based on the status of nutritional intake, Gastroenterology Research and Practice, 10.1155/2012/859697, 2012.12, The dietary intake of patients with nonalcoholic fatty liver disease (NAFLD) is generally characterized by high levels of carbohydrate, fat, and/or cholesterol, and these dietary patterns influence hepatic lipid metabolism in the patients. Therefore, careful investigation of dietary habits could lead to better nutrition therapy in NAFLD patients. The main treatment for chronic hepatitis C (CHC) is interferon-based antiviral therapy, which often causes a decrease in appetite and energy intake; hence, nutritional support is also required during therapy to prevent undernourishment, treatment interruption, and a reduction in quality of life. Moreover, addition of some nutrients that act to suppress viral proliferation is recommended. As a substitutive treatment, low-iron diet therapy, which is relatively safe and effective for preventing hepatocellular carcinoma, is also recommended for CHC patients. Some patients with liver cirrhosis (LC) have decreased dietary energy and protein intake, while the number of LC patients with overeating and obesity is increasing, indicating that the nutritional state of LC patients has a broad spectrum. Therefore, nutrition therapy for LC patients should be planned on an assessment of their complications, nutritional state, and dietary intake. Late evening snacks, branched-chain amino acids, zinc, and probiotics are considered for effective nutritional utilization..
47. Kenichiro Yasutake, Machiko Bekki, Masako Ichinose, Michiko Ikemoto, Tatsuya Fujino, Tomoki Ryu, Yoshiyuki Wada, Yuko Takami, Hideki Saitsu, Motoyuki Kohjima, Kunitaka Fukuizumi, Manabu Nakashima, Makoto Nakamuta, Munechika Enjoji, Assessing current nutritional status of patients with HCV-related liver cirrhosis in the compensated stage, Asia Pacific Journal of Clinical Nutrition, 21, 3, 400-405, 2012.09, Background/Aim: Nutritional states of Japanese patients with liver cirrhosis have recently shown great diversity, some show protein energy malnutrition and others excessive nutrition and obesity. For there to be adequate guidance regarding dietary treatment, it is important that a patient's current nutritional state be clarified. Methods: We assessed nutritive intake in Japanese cirrhotic patients and determined their nutritional problems. Subjects were non-hospitalized patients with hepatitis C virus (HCV)-related cirrhosis in the compensated stage (n=47), chronic hepatitis C (n=46) or healthy volunteers (n=32). A brief self-administered diet history questionnaire was conducted with assistance from a registered dietitian. Results: We categorized patients with cirrhosis according to daily intake of energy and protein; 10.6% had an energy and protein intake within a normal range, 72.4% showed excessive intake, and 17.0% showed insufficient intake of energy or protein. In cirrhotic patients with diabetic complications, the intake levels of energy, proteins, fat and carbohydrates were significantly higher than in patients without diabetes. Moreover, cirrhotic patients had significantly higher intake levels of energy, protein and fat than did chronic hepatitis C patients and healthy individuals. In patients with HCV-related liver cirrhosis, nsufficient intake of energy and protein was shown in some, while many, especially tihose with diabetes, showed excessive intake. Conclusion: For nutritive management of cirrhotic patients, the intake of various nutrients should be appropriately assessed and effective nutritional education systems established..
48. Masayuki Miyazaki, Masaki Kato, Masatake Tanaka, Kosuke Tanaka, Shinichiro Takao, Motoyuki Kohjima, Tetsuhide Ito, Munechika Enjoji, Makoto Nakamuta, Kazuhiro Kotoh, Ryoichi Takayanagi, Antithrombin III injection via the portal vein suppresses liver damage, World Journal of Gastroenterology, 10.3748/wjg.v18.i16.1884, 18, 16, 1884-1891, 2012.04, AIM: To investigate the effects of antithrombin III (AT III) injection via the portal vein in acute liver failure. METHODS: Thirty rats were intraperitoneally challenged with lipopolysaccharide (LPS) and D-galactosamine (GalN) and divided into three groups: A control group; a group injected with AT III via the tail vein; and a group injected with AT III via the portal vein. AT III (50 U/kg body weight) was administrated 1 h after challenge with LPS and GalN. Serum levels of inflammatory cytokines and fibrin degradation products, hepatic fibrin deposition, and hepatic mRNA expression of hypoxiarelated hypoxiarelated genes were analyzed. RESULTS: Serum levels of alanine aminotransferase, tumor necrosis factor-α and interleukin-6 decreased significantly following portal vein AT III injection compared with tail vein injection, and control rats. Portal vein AT III injection reduced liver cell destruction and decreased hepatic fibrin deposition. This treatment also significantly reduced hepatic mRNA expression of lactate dehydrogenase and heme oxygenase-1. CONCLUSION: A clinically acceptable dose of AT III injection into the portal vein suppressed liver damage, probably through its enhanced anticoagulant and antiinflammatory activities..
49. Makoto Nakamuta, Motoyuki Kohjima, Lipid metabolism-related gene expression in hepatocellular carcinoma, Journal of Japanese Society of Gastroenterology, 109, 4, 555-562, 2012.04.
50. Masayuki Miyazaki, Masaki Kato, Kosuke Tanaka, Masatake Tanaka, Motoyuki Kohjima, Kazuhiko Nakamura, Munechika Enjoji, Makoto Nakamuta, Kazuhiro Kotoh, Ryoichi Takayanagi, Increased hepatic expression of dipeptidyl peptidase-4 in non-alcoholic fatty liver disease and its association with insulin resistance and glucose metabolism, Molecular medicine reports, 10.3892/mmr.2011.707, 5, 3, 729-733, 2012.03, Dipeptidyl peptidase-4 (DPP4) is a serine protease that degrades glucagon-like peptide-1 (GLP-1), an incretin hormone that stimulates insulin secretion from pancreatic β-cells. DPP4 is also involved in the regulation of T cell-mediated inflammatory processes. These properties of DPP4 suggest that it may play a role in the progression of non-alcoholic fatty liver disease (NAFLD). Hepatic DPP4 mRNA expression levels were analyzed by real-time PCR using liver biopsy samples from 17 NAFLD patients and 10 healthy subjects. In NAFLD patients, we also examined correlations between DPP4 expression levels and metabolic factors, including homeostasis model assessment-insulin resistance (HOMA-IR), body mass index (BMI), and serum cholesterol and triglyceride levels. To examine the potential effects of nutritional factors, DPP4 expression levels were analyzed in HepG2 cells subjected to various culture conditions. Hepatic DPP4 mRNA expression was significantly greater in NAFLD patients than in control subjects. DPP4 expression levels were negatively correlated with HOMA-IR and positively correlated with serum cholesterol levels. In HepG2 cells, high glucose significantly enhanced DPP4 expression, whereas insulin, fatty acids and cholesterol did not. Increased hepatic expression of DPP4 in NAFLD may be associated with metabolic factors, including insulin resistance, and may adversely affect glucose metabolism in this liver disease..
51. Kazuhiro Kotoh, Marie Fukushima, Yuki Horikawa, Shinsaku Yamashita, Motoyuki Kohjima, Makoto Nakamuta, Munechika Enjoji, Serum albumin is present at higher levels in alcoholic liver cirrhosis as compared to HCV-related cirrhosis, Experimental and Therapeutic Medicine, 10.3892/etm.2011.370, 3, 1, 72-75, 2012.01, Residual hepatic functional reserve in cirrhotic patients is generally evaluated by a multivariate scoring system (Child-Pugh classification), which includes serum albumin levels as a variable. However, several patients show discrepancies between serum albumin levels and the progression of liver fibrosis, especially those with alcoholic cirrhosis. To assess whether hepatic capacity of protein synthesis varies with the etiology of cirrhosis, serum albumin and cholinesterase levels, and prothrombin time were compared between alcoholic cirrhosis and hepatitis C virus (HCV)-related cirrhosis. To minimize the influence of malnutrition and extrahepatic platelet destruction, patients with hepatocellular carcinoma, uncontrolled diabetes, appetite loss and/or splenal longitudinal size >15 cm were excluded. The patients with compensated liver cirrhosis were divided into three groups as follows: alcohol +/HCV + (alcohol + HCV group; n=31), alcohol -/HCV + (HCV group; n=31) and alcohol +/HCV - (alcohol group; n=27). These groups were adjusted with respect to age, gender, body mass index and platelet count. Serum albumin levels in the alcohol group were significantly higher than those in the HCV group, with a difference of approximately 0.5 g/dl in every class of platelet count. The correlation of the alcohol + HCV group was intermediate between the alcohol and HCV groups. On the other hand, the correlations between serum cholinesterase levels and platelet counts were similar among the three groups. The prothrombin time was also comparable among the groups. Accordingly, serum albumin levels were higher in patients with alcoholic cirrhosis and alcohol consumption should be carefully considered when evaluating hepatic functional reserve..
52. Nobito Higuchi, Masaki Kato, Masatake Tanaka, Masayuki Miyazaki, Shinichiro Takao, Motoyuki Kohjima, Kazuhiro Kotoh, Munechika Enjoji, Makoto Nakamuta, Ryoichi Takayanagi, Effects of insulin resistance and hepatic lipid accumulation on hepatic mRNA expression levels of apoB, MTP and L-FABP in non-alcoholic fatty liver disease, Experimental and Therapeutic Medicine, 10.3892/etm.2011.328, 2, 6, 1077-1081, 2011.11, Non-alcoholic fatty liver disease (NAFLD) is considered a hepatic manifestation of metabolic syndrome, which is known to be associated with insulin resistance (IR). NAFLD occurs when the rate of hepatic fatty acid uptake from plasma and de novo fatty acid synthesis is greater than the rate of fatty acid oxidation and excretion as very low-density lipoprotein (VLDL). To estimate the effects of IR on hepatic lipid excretion, mRNA expression levels of genes involved in VLDL assembly were analyzed in NAFLD liver. Twenty-two histologically proven NAFLD patients and 10 healthy control subjects were enrolled in this study. mRNA was extracted from liver biopsy samples and real-time PCR was performed to quantify the expression levels of apolipoprotein B (apoB), microsomal triglyceride transfer protein (MTP) and liver fatty-acid binding protein (L-FABP). Hepatic expression levels of the genes were compared between NAFLD patients and control subjects. In NAFLD patients, we also examined correlations between expression levels of the genes and metabolic factors, including IR, and the extent of obesity and hepatic lipid accumulation. Hepatic expression levels of apoB, MTP and L-FABP were significantly up-regulated in NAFLD patients compared to control subjects. The expression levels of MTP were correlated with those of apoB, but not with those of L-FABP. In the NAFLD liver, the expression levels of MTP were significantly reduced in patients with HOMA-IR >2.5. In addition, a significant reduction in MTP expression was observed in livers with advanced steatosis. Enhanced expression of genes involved in VLDL assembly may be promoted to release excess lipid from NAFLD livers. However, the progression of IR and hepatic steatosis may attenuate this compensatory process..
53. Mako Toyoda, Akira Kitaoka, Kazuyuki Machida, Takuya Nishinakagawa, Ryoko Yada, Motoyuki Kohjima, Masaki Kato, Kazuhiro Kotoh, Naoya Sakamoto, Goshi Shiota, Makoto Nakamuta, Manabu Nakashima, Munechika Enjoji, Association between lipid accumulation and the cannabinoid system in Huh7 cells expressing HCV genes, International journal of molecular medicine, 10.3892/ijmm.2011.622, 27, 5, 619-624, 2011.05, Evidence from clinical and laboratory studies has accumulated indicating that the activation of the cannabinoid system is crucial for steatosis, especially in non-alcoholic fatty liver disease. However, the association between hepatitis C C virus (HCV) infection and the cannabinoid system has not been well investigated and it is unclear whether steatosis in chronic hepatitis C develops via activation of the endo-cannabinoid/cannabinoid receptor signaling pathway. In this study, we examined the expression of a cannabinoid receptor (CB1) and the lipid accumulation in the hepatic Huh7 cell line, expressing HCV genes. We utilized Huh7/Rep-Feo-1b cells stably expressing HCV non-structural proteins (NS) 3, NS4, NS5A, and NS5B, as well as Tet-On Core-2 cells, in which the HCV core protein expression is inducible. Significantly higher levels of stored triglycerides were found in Huh7/Rep-Feo-1b cells compared to Huh7 cells. Also, triglyceride accumulation and CB1 receptor expression were down-regulated in Huh7/Rep-Feo-1b cells after HCV reduction by IFNα. Moreover, lipid accumulation appeared to increase after CB1 agonist treatment, while it decreased after CB1 antagonist treatment, although significant differences were not found compared to untreated cells. In Tet-On Core-2 cells, induction of HCV core protein expression did not affect CB1 expression or triglyceride accumulation. The results of this study in cultured cells suggest that HCV infection may activate the cannabinoid system and precede steatosis, but the core protein by itself may not have any effect on the cannabinoid system..
54. M. Nakamuta, T. Fujino, R. Yada, Y. Aoyagi, K. Yasutake, M. Kohjima, K. Fukuizumi, T. Yoshimoto, N. Harada, M. Yada, M. Kato, K. Kotoh, A. Taketomi, Y. Maehara, M. Nakashima, M. Enjoji, Expression profiles of genes associated with viral entry in HCV-infected human liver, Journal of Medical Virology, 10.1002/jmv.22042, 83, 5, 921-927, 2011.05, Recent studies have demonstrated that several cellular factors are involved in entry of hepatitis C virus (HCV) into host cells. Detailed gene expression profiles of these factors in HCV-infected livers have not been reported for humans. Transcriptional levels of LDL receptor (LDLR), CD81, scavenger receptor class B type I (SR-BI), claudin-1, and occludin genes in liver samples from patients with chronic hepatitis C were investigated. Serum levels of LDL-cholesterol (LDL-C) and HCV core antigen were also evaluated, and expression of claudin-1 and occludin were immunohistochemically analyzed. Compared with normal liver, transcription of LDLR and claudin-1 genes was significantly suppressed (P<0.0001) and occludin transcription was significantly up-regulated in HCV-infected livers (P<0.0001). Significant positive correlations were found for LDLR versus occludin, LDLR versus claudin-1, occludin versus claudin-1, and CD81 versus SR-BI in HCV-infected (P=0.0012, P<0.0001, P=0.0004, and P<0.0001, respectively) and normal livers (P<0.0001, P=0.0051, P<0.0001, and P<0.0001, respectively). Positive correlation was observed between serum levels of HCV core antigen and LDL-C (P=0.0147), with their levels negatively correlated to LDLR (P=0.0270 and P=0.0021, respectively). Immunohistochemically, hepatocellular expression of claudin-1 and occludin was increased in HCV-infected livers. Different levels of expression were demonstrated at the mRNA and protein levels for occludin and claudin-1 in HCV-infected and normal livers. Correlation of elements associated with viral entry was comparable in HCV-infected and normal livers..
55. Kazuhiro Kotoh, Masaki Kato, Motoyuki Kohjima, Masatake Tanaka, Masayuki Miyazaki, Kazuhiko Nakamura, Munechika Enjoji, Makoto Nakamuta, Ryoichi Takayanagi, Lactate dehydrogenase production in hepatocytes is increased at an early stage of acute liver failure, Experimental and Therapeutic Medicine, 10.3892/etm.2011.197, 2, 2, 195-199, 2011.03, Although the mechanism involved in acute liver failure (ALF) has not yet been clarified, microcirculatory disturbance in the liver appears to play a pivotal role in the progression of this disease. To confirm the existence of hepatic hypoxic conditions, we evaluated the amounts of lactate dehydrogenase (LDH) in hepatocytes, since its production increases under low oxygen concentrations. Histological examination was performed in 7 patients with ALF. All 7 patients underwent a liver biopsy during the acute phase of ALF, and 4 of them underwent a second biopsy during the recovery phase. The obtained samples were immunohistochemically stained with anti-LDH5 and anti-CD-68 antibodies. As controls, we examined samples from patients with acute hepatitis, chronic hepatitis and liver cirrhosis. The production of LDH by hepatocytes and the number of CD-68 positive macrophages were markedly increased at the acute phase of ALF, and both of these effects abruptly decreased during the recovery phase. By contrast, most of the samples from the patients with chronic hepatitis and acute hepatitis showed slightly any increase in LDH staining. In cirrhotic patients, partially elevated LDH production was observed mainly around the central vein, but the staining intensity was less compared to that in ALF patients. Our findings indicate that hepatic hypoxic conditions exist in ALF at the acute phase and seem to closely correlate with macrophage overactivation in the liver. We speculate that microcirculatory disturbance may be a key process in the development and progression of ALF..
56. Masayuki Miyazaki, Masaki Kato, Masatake Tanaka, Kosuke Tanaka, Shinichiro Takao, Motoyuki Kohjima, Tetsuhide Ito, Munechika Enjoji, Makoto Nakamuta, Kazuhiro Kotoh, Ryoichi Takayanagi, Contrast-enhanced ultrasonography using Sonazoid to evaluate changes in hepatic hemodynamics in acute liver injury, Journal of Gastroenterology and Hepatology (Australia), 10.1111/j.1440-1746.2011.06790.x, 26, 12, 1749-1756, 2011.01, Background and Aims: Disturbances in hepatic microcirculation are believed to be involved in the mechanisms regulating the progression of acute liver injury (ALI). Evaluation of hepatic hemodynamics in patients with acute liver injury might be helpful in understanding the extent of the intrahepatic microcirculatory disturbances. Therefore, we investigated whether contrast-enhanced ultrasonography (CEUS) is useful to evaluate the changes in hepatic hemodynamics in patients with ALI. Methods: CEUS was performed in 21 patients with ALI and coagulopathy. Participants were injected with 0.0075mL Sonazoid/kg body weight, and time-intensity curves were simultaneously recorded for the hepatic and portal veins. The data were compared with those of 10 healthy volunteers. Results: The arrival time of Sonazoid in the hepatic vein was similar to that in the portal vein in the patients, whereas the arrival time in the hepatic vein was delayed relative to that in the portal vain in the controls (interval between the hepatic and portal vein arrival times, control vs patients 6.74±3.07s vs 1.13±1.07s, P<0.001). Repeated examination revealed that the interval between the hepatic and portal vein arrival times was extended by improvements in hepatic function. The early arrival of Sonazoid in the hepatic vein in the patients is likely to reflect the formation of intrahepatic shunts as a result of hepatic microcirculatory disturbances. Conclusion: CEUS using Sonazoid is a useful method to estimate the changes in hepatic hemodynamics in patients with ALI..
57. Tatsuya Fujino, Makoto Nakamuta, Yoko Aoyagi, Motoyuki Kohjima, Takeaki Satoh, Mika Fukuda, Hiromi Ishibashi, Hiroshi Yatsuhashi, Munechika Enjoji, Early dynamics of viremia in patients with genotype 1b chronic hepatitis C
Peg-IFNα2a shows earlier viral decline than peg-IFNα2b in combination therapy with ribavirin, Medical Science Monitor, 10.12659/MSM.882127, 17, 12, CR687-CR691, 2011.01, Background: We aimed to assess differences in early viral dynamics following treatment with either peg-IFNα2a or peg-IFNα2b in combination with ribavirin in patients with chronic genotype 1b HCV infection. Material/Methods: Sixty-one patients in the peg-IFNα2a + ribavirin treatment (group α2a) and 88 patients in the peg- IFNα2b + ribavirin treatment (group α2b) were retrospectively analyzed. The early dynamics of HCV RNA over 12 weeks were evaluated. Sustained virological response (SVR) was defined as undetectable HCV RNA at week 24 after end of therapy. First- (day 0-1) and second-phase (day 1-28) viral decline rates were calculated in accordance with theoretical formulae. Results: Baseline HCV RNA concentrations were almost similar between the 2 groups. In group α2a, viral decline was significantly greater than in group α2b at weeks 4, 8, and 12. In group α2a, viral decline was significantly greater in SVR patients than in non-SVR patients at week 2, whereas significantly greater viral decline in SVR patients was found during weeks 1-12 in group α2b. The first-phase viral decline rate was significantly larger in group α2a than in group α2b (1.31±0.84 vs. 0.70±0.97 log IU/mL/day; p<0.0001). Within SVR patients, first-phase viral decline rate was significantly larger in group α2a compared with group α2b (1.45±0.85 vs. 0.78±1.0 log IU/mL/day; p<0.0001). Second-phase viral decline rate was comparable between the groups. Conclusions: Peg-IFNα2a showed earlier viral decline than peg-IFNα2b and the difference was obvious, especially in the first-phase viral decline..
58. Nobito Higuchi, Masaki Kato, Masayuki Miyazaki, Masatake Tanaka, Motoyuki Kohjima, Tetsuhide Ito, Makoto Nakamuta, Munechika Enjoji, Kazuhiro Kotoh, Ryoichi Takayanagi, Potential role of Branched-chain amino acids in glucose metabolism through the accelerated induction of the glucose-sensing apparatus in the liver, Journal of Cellular Biochemistry, 10.1002/jcb.22688, 112, 1, 30-38, 2011.01, Branched-chain amino acids (BCAAs) have a potential to improve glucose metabolism in cirrhotic patients; however, the contribution of liver in this process has not been clarified. To estimate the effect of BCAA on glucose metabolism in liver, we evaluated the mRNA expression levels of glucose-sensing apparatus genes in HepG2 cells and in rat liver after oral administration of BCAA. HepG2 cells were cultured in low glucose (100 mg/dl) or high glucose (400 mg/dl) in the absence or presence of BCAA. The mRNA expression levels and protein levels of GLUT2 and liver-type glucokinase (L-GK) were estimated using RT-PCR and immunoblotting. The expression levels of transcriptional factors, including SREBP-1c, ChREBP, PPAR-gm and LXRα, were estimated. The mRNA expression levels of transcriptional factors, glycogen synthase, and genes involved in gluconeogenesis were evaluated in rat liver at 3 h after the administration of BCAA. BCAA accelerated the expression of GLUT2 and L-GK in HepG2 cells in high glucose. Expression levels of ChREBP, SREBP-1c, and LXRα were also increased in this condition. BCAA administration enhanced the mRNA expression levels of L-GK, SREBP-1c, and LXRα and suppressed the expression levels of G-6-Pase in rat liver, without affecting the expression levels of glycogen synthase or serum glucose concentrations. BCAA administration enhanced the bioactivity of the glucose-sensing apparatus, probably via the activation of a transcriptional mechanism, suggesting that these amino acids may improve glucose metabolism through the accelerated utility of glucose and glucose-6-phosphate in the liver. J. Cell..
59. Kazuhiro Kotoh, Masaki Kato, Motoyuki Kohjima, Makoto Nakamuta, Munechika Enjoji, A new treatment strategy for acute liver failure, World Journal of Hepatology, 10.4254/wjh.v2.i11.395, 2, 11, 395-400, 2010.12, Acute liver failure (ALF) is a syndrome defined by coa gulopathy and encephalopathy and no effective treatments have been established, except for liver transplantation. However, considering the limited supply of donors, we should endeavor to prevent the progression of this syndrome in its early stage to improve the prognosis of patients with ALF. Recently, several authors have reported that over-activation of intrahepatic macrophages plays an important role in the progression of ALF and we have developed a new treatment method, transcatheter arterial steroid injection therapy (TASIT), to suppress macrophage activation. We have now used TASIT for 5 years and have found that TASIT is effective for patients with over-activation of macrophages in the liver but not for those with lesser activation of macrophages. Therefore, to identify the most appropriate patients for TASIT, we tried to categorize patients with ALF or acute liver injury according to markers for the degree of intrahepatic macrophage activation. This approach was helpful to select the appropriate treatment including liver transplantation. We believe that it is essential to analyze disease progression in each patient before selecting the most appropriate treatment..
60. Munechika Enjoji, Kazuyuki MacHida, Motoyuki Kohjima, Masaki Kato, Kazuhiro Kotoh, Kazuhisa Matsunaga, Manabu Nakashima, Makoto Nakamuta, NPC1L1 inhibitor ezetimibe is a reliable therapeutic agent for non-obese patients with nonalcoholic fatty liver disease, Lipids in Health and Disease, 10.1186/1476-511X-9-29, 9, 2010.12, Background: We recently examined the distribution of abdominal fat, dietary intake and biochemical data in patients with nonalcoholic fatty liver disease (NAFLD) and found that non-obese NAFLD patients did not necessarily exhibit insulin resistance and/or dysregulated secretion of adipocytokines. However, dietary cholesterol intake was superabundant in non-obese patients compared with obese patients, although total energy and carbohydrate intake was not excessive. Therefore, excess cholesterol intake appears to be one of the main factors associated with NAFLD development and liver injury. Methods. We reviewed a year of follow-up data of non-obese NAFLD patients treated with Niemann-Pick C1 like 1 inhibitor ezetimibe to evaluate its therapeutic effect on clinical parameters related to NAFLD. Without any dietary or exercise modification, 10 mg/day of ezetimibe was given to 8 patients. In 4 of 8 patients, ezetimibe was administered initially. In the remaining 4 patients, medication was switched from ursodeoxycholic acid to ezetimibe. Results: In each patient, body mass index was maintained under 25 kg/m2 during the observation period. Serum ALT levels significantly decreased within 6 months and in 4 patients levels reached the normal range (<30 U/L), which was accompanied with at least a 10% decrease in serum total cholesterol and LDL-cholesterol. However, ultrasonographic findings of fatty liver did not show obvious improvement for a year. Conclusion: We conclude that the cholesterol absorption inhibitor ezetimibe can suppress hepatic injury in non-obese patients with NAFLD and that ezetimibe may offer a novel treatment for NAFLD..
61. Tatsuya Fujino, Makoto Nakamuta, Ryoko Yada, Yoko Aoyagi, Kenichiro Yasutake, Motoyuki Kohjima, Kunitaka Fukuizumi, Tsuyoshi Yoshimoto, Naohiko Harada, Masayoshi Yada, Masaki Kato, Kazuhiro Kotoh, Akinobu Taketomi, Yoshihiko Maehara, Manabu Nakashima, Munechika Enjoji, Expression profile of lipid metabolism-associated genes in hepatitis C virus-infected human liver, Hepatology Research, 10.1111/j.1872-034X.2010.00700.x, 40, 9, 923-929, 2010.09, Aim: Recent studies have shown that lipid metabolic pathways are required for the entry, replication and secretion of hepatitis C virus (HCV). Although little is known about the life cycle of HCV in humans, the activation of cholesterol and fatty acid biosynthesis may be critical for HCV proliferation. Methods: We assessed the transcription levels of genes essential for cholesterol and fatty acid biosynthesis in liver samples obtained from patients with chronic hepatitis C and determined their correlations. The serum levels of low-density lipoprotein (LDL) cholesterol and HCV core antigen were also measured. Results: The gene expression of the LDL receptor (LDLR) was suppressed, whereas that of SREBP1c, liver X receptor-α (LXRα), fatty acid synthase (FASN), and HMG-CoA reductase and synthase (HMGR and HMGS) was significantly increased, and SREBP2 transcription was comparable in HCV-infected liver compared with normal liver. Positive correlations were found for LDLR versus HMGR, HMGR versus SREBP1c, and LDLR versus SREBP2 in the HCV-infected and control liver. Although the LXRα-SREBP1c-FASN pathway was upregulated, proteasome activator 28γ (PA28γ) was downregulated at the transcriptional level in HCV-infected liver, and was not significantly correlated with the other genes examined. The serum LDL cholesterol level was negatively correlated with LDLR and HMGR expression. Conclusion: These results suggest that, in HCV-infected liver, the cholesterol load increases and cholesterol uptake is controlled, while de novo cholesterol synthesis is upregulated compared with the normal physiological state. The positive correlations in the expression levels of some cholesterol metabolism-associated genes indicate that not all of the metabolic pathways are dysregulated in HCV-infected liver..
62. Motoyuki Kohjima, Yuxiang Sun, Lawrence Chan, Increased food intake leads to obesity and insulin resistance in the Tg2576 Alzheimer's disease mouse model, Endocrinology, 10.1210/en.2009-1196, 151, 4, 1532-1540, 2010.04, Recent studies suggest that hyperinsulinemia and insulin resistance are linked to Alzheimer's disease (AD). In this study, we used Tg2576 transgenic (Tg) mice, a widely used transgenic mouse model for AD, to explore the relationship between increased amyloid β-peptide (Aβ) and insulin resistance. When fed a high-fat diet (HFD), Tg mice developed obesity and insulin resistance at 16 wk of age. Furthermore, HFD-fed Tg mice displayed abnormal feeding behavior and increased caloric intake with time. Although caloric intake of HFD-fed Tg mice was similar to that of normal diet-fed Tg or wild-type mice during 4 to 8 wk of age, it increased sharply at 12 wk, and went up further at 16 wk, which paralleled changes in the level of Aβ40 and Aβ42 in the brain of these mice. Limiting food intake in HFD-fed Tg mice by pair-feeding a caloric intake identical with that of normal diet-fed mice completely prevented the obesity and insulin intolerance of HFD-fed Tg mice. The hypothalamus of HFD-fed Tg mice had a significant decrease in the expression of the anorexigenic neuropeptide, brain-derived neurotrophic factor, at both the mRNA and protein levels. These findings suggest that the increased Aβ in the brain of HFD-fed Tg2576 mice is associated with reduced brain-derived neurotrophic factor expression, which led to abnormal feeding behavior and increased food intake, resulting in obesity and insulin resistance in these animals..
63. M. Nakamuta, T. Fujino, R. Yada, K. Yasutake, T. Yoshimoto, N. Harada, M. Yada, N. Higuchi, M. Kato, M. Kohjima, A. Taketomi, Y. Maehara, T. Nishinakagawa, K. Machida, K. Matsunaga, M. Nakashima, K. Kotoh, M. Enjoji, Therapeutic effect of bezafibrate against biliary damage
A study of phospholipid secretion via the PPARα-MDR3 pathway, International Journal of Clinical Pharmacology and Therapeutics, 48, 1, 22-28, 2010.01, Objective: Bezafibrate (BF) has been used to treat biliary damage, particularly in patients with primary biliary cirrhosis (PBC), and its clinical efficacy has been demonstrated. The mechanism of action is thought to involve activation of the PPARα-MDR3-phospholipid (PL) secretion pathway. We tried to confirm this hypothesis in patients with hepatobiliary disease. Methods: The levels of serum -glutamyl transpeptidase and alkaline phosphatase, and those of bile components were examined before and after BF administration in patients with obstructive jaundice undergoing percutaneous transhepatic biliary drainage (PTBD). Hepatic expression of PPARα and MDR3 was quantified by real-time PCR in patients with PBC or non-alcoholic fatty liver disease (NAFLD). Results: In patients with obstructive jaundice, BF decreased the serum levels of biliary enzymes and increased the bile concentration of PL. In patients with PBC or NAFLD, the expression levels of MDR3 were already up-regulated before starting the BF treatment. Although BF treatment did not further up-regulate MDR3 expression in NAFLD patients, PPARα expression was significantly increased. Conclusions: BF enhanced the secretion of PL into bile in cholestatic patients undergoing PTBD. However, in patients with PBC or NAFLD, diseases that represent cholesterol overload, MDR3 was already expressed at a high level to compensate for bile acids overproduction, and its expression was hardly affected by BF. In patients with chronic liver diseases such as PBC, BF may induce clinical effects via mechanisms independent of PL secretion..
64. Makoto Nakamuta, Ryoko Yada, Tatsuya Fujino, Masayoshi Yada, Nobito Higuchi, Masatake Tanaka, Masayuki Miyazaki, Motoyuki Kohjima, Masaki Kato, Tsuyoshi Yoshimoto, Naohiko Harada, Akinobu Taketomi, Yoshihiko Maehara, Momoko Koga, Takuya Nishinakagawa, Manabu Nakashima, Kazuhiro Kotoh, Munechika Enjoji, Changes in the expression of cholesterol metabolism-associated genes in HCV-infected liver
A novel target for therapy?, International journal of molecular medicine, 10.3892/ijmm_00000299, 24, 6, 825-828, 2009.11, Recent investigations indicate that hepatitis C virus (HCV) infection is closely associated with hepatocytic lipid metabolism and induces hepatic steatosis. However, the actual lipid metabolism in HCV-infected liver has not been extensively investigated in humans. In this study, we evaluated the expression of lipid metabolism-associated genes in patients with HCV infection by real-time PCR. Sterol regulatory element-binding protein (SREBP)-2 expression was unchanged and low density lipoprotein receptor expression was markedly reduced by 90% in HCV-infected liver. The expression of apolipoprotein B100, microsomal triglyceride transfer ptotein and ATP-binding cassette G5 was significantly increased. Up-regulation of cholesterol synthesis-associated genes, including HMG-CoA reductase, HMG-CoA synthase, farnesyl-diphosphate synthase and squalene synthase, confirmed enhanced de novo cholesterol synthesis. The expression of cholesterol 7α-hydroxylase and farnesoid X receptor was enhanced, while bile salt export pump expression was unchanged. Fatty acid synthase expression was increased which was accompanied by increased expression of liver X receptor α and SREBP-1c. In summary, the regulation of lipid metabolism was impaired and cholesterol and fatty acid synthesis continued to increase without negative feedback in HCV-infected liver. These changes may be beneficial for HCV replication..
65. S. L. Samson, M. Kohjima, L. Chan, Autoregulatory gene therapy for obesity
A fine balance: An autoregulatory gene therapy approach to treat obesity and achieve energy homeostasis, Gene Therapy, 10.1038/gt.2009.88, 16, 10, 1175-1177, 2009.07.
66. Makoto Nakamuta, Tatsuya Fujino, Ryoko Yada, Masayoshi Yada, Kenichiro Yasutake, Tsuyoshi Yoshimoto, Naohiko Harada, Nobito Higuchi, Masaki Kato, Motoyuki Kohjima, Akinobu Taketomi, Yoshihiko Maehara, Manabu Nakashima, Kazuhiro Kotoh, Munechika Enjoji, Impact of cholesterol metabolism and the LXRα-SREBP-1c pathway on nonalcoholic fatty liver disease, International journal of molecular medicine, 10.3892/ijmm_00000170, 23, 5, 603-608, 2009.07, We previously studied fatty acid metabolism in the liver of nonalcoholic fatty liver disease (NAFLD) and reported the activation of the LXRα-SREBP-1c pathway in hepatocytes. LXRα regulates cholesterol metabolism as well as fatty acid metabolism, and its agonistic ligands are oxysterols. Moreover, there is some evidence that excess cholesterol intake is involved in the onset of NAFLD. Therefore, in this study, we examined the expression of cholesterol metabolism-associated genes in the NAFLD liver by real-time PCR. Expression of LXRα and ACAT1 was up-regulated in NAFLD and this was more noticeable in non-obese rather than in obese patients. Although the expression of the LDL receptor, which acts on cholesterol uptake, and of SREBP-2, a positive key regulator of cholesterol, was suppressed, the expression of enzymes that promote cholesterol synthesis was uniformly increased in NAFLD. Gene expression of apoB100 and microsomal triglyceride transfer protein, which are associated with VLDL secretion, and ABCG5, which is involved in cholesterol excretion, was significantly elevated in NAFLD. Because cholesterol accumulates in hepatocytes in NAFLD liver, cholesterol uptake and synthesis should be physiologically down-regulated. However, cholesterol synthesis was activated in NAFLD liver, meaning that cholesterol metabolism is dysregulated in NAFLD. Over-production of cholesterol may lead to an increased level of oxysterols, activation of LXRα and SREBP-1c, and enhanced fatty acid synthesis..
67. Munechika Enjoji, Ryoko Yada, Tatsuya Fujino, Tsuyoshi Yoshimoto, Masayoshi Yada, Naohiko Harada, Nobito Higuchi, Masaki Kato, Motoyuki Kohjima, Akinobu Taketomi, Yoshihiko Maehara, Manabu Nakashima, Kazuhiro Kotoh, Makoto Nakamuta, The state of cholesterol metabolism in the liver of patients with primary biliary cirrhosis
The role of MDR3 expression, Hepatology International, 10.1007/s12072-009-9137-y, 3, 3, 490-496, 2009.06, Aim: Because dyslipidemia, such as hypercholesterolemia, is a characteristic of primary biliary cirrhosis (PBC), hepatic lipid metabolism may be disturbed in PBC patients. We examined the expression of lipid metabolism-associated genes in PBC liver. Methods: All of the patients examined were in stage I or II PBC and without medication. RNA was isolated from liver specimens by needle biopsies of PBC patients and controls. The expression levels of various genes were measured by real-time RT-PCR. Multidrug resistance 3 (MDR3) expression was examined immunohistochemically. Statistical correlations between the gene expression levels and indices of blood testing were calculated. Results: The expression levels of sterol regulatory element-binding protein (SREBP) 2 and LDL receptor were significantly lower, and those of apolipoprotein B, microsomal triglyceride transfer protein, ATP-binding cassette G5, and liver X receptor α (LXRα) were significantly higher in the PBC liver than in the normal control liver. The expression levels of bile acid synthesis- and excretion-associated genes did not change, and those of farnesoid X receptor, peroxisome proliferator-activated receptor α, and SREBP-1c were similar between the PBC and normal liver. MDR3 gene expression levels in the PBC liver were more than 4-fold higher than those in the control liver. Immunohistochemically, strong canalicular staining for MDR3 was observed in the PBC liver. LXRα expression was positively correlated with MDR3 levels. Serum levels of α-glutamyl transpeptidase (GGT) and IgM were negatively correlated with MDR3 levels. Conclusions: Hepatocellular cholesterol metabolism was at least partially disturbed, even in the early stage of PBC. The most characteristic finding was a distinct elevation of MDR3 expression, and the MDR3 levels were negatively correlated with GGT and IgM levels..
68. Motoyuki Kohjima, Munechika Enjoji, Nobito Higuchi, Masaki Kato, Kazuhiro Kotoh, Manabu Nakashima, Makoto Nakamuta, The effects of unsaturated fatty acids on lipid metabolism in HepG2 cells, In Vitro Cellular and Developmental Biology - Animal, 10.1007/s11626-008-9144-7, 45, 1-2, 6-9, 2009.02, We investigated the effects of stearic acid (saturated), oleic acid (monounsaturated), linoleic acid (n-6 polyunsaturated), and α-linolenic acid (n-3 polyunsaturated) on lipid metabolism in a hepatocyte-derived cell line, HepG2. HepG2 cells were cultured in medium supplemented with either stearic acid (0.1% w/v), oleic acid (0.1% v/v), linoleic acid (0.1% v/v), or α-linolenic acid (0.1% v/v). After 24 h, expression of lipid metabolism-associated genes was evaluated by real-time PCR. α-Linolenic acid showed a suppressive effect on the hepatic fatty acid de novo synthesis and fatty acid oxidation pathways, while linoleic acid also showed a tendency to suppress these pathways although the effect was weaker. Moreover, α-linolenic acid enhanced the expression of enzymes associated with reactive oxygen species (ROS) elimination. In contrast, oleic acid tended to promote fatty acid synthesis and oxidation. In conclusion, α-linolenic acid and linoleic acid may be expected to ameliorate hepatic steatosis by downregulating fatty acid de novo synthesis and fatty acid oxidation, and by upregulating ROS elimination enzymes. Oleic acid had no distinct effects for improving steatosis or oxidative stress..
69. Makoto Nakamuta, Motoyuki Kohjima, Nobito Higuchi, Masaki Kato, Kazuhiro Kotoh, Tsuyoshi Yoshimoto, Masayoshi Yada, Ryoko Yada, Ryosuke Takemoto, Kunitaka Fukuizumi, Naohiko Harada, Akinobu Taketomi, Yoshihiko Maehara, Manabu Nakashima, Munechika Enjoji, The significance of differences in fatty acid metabolism between obese and non-obese patients with non-alcoholic fatty liver disease, International journal of molecular medicine, 10.3892/ijmm_00000070, 22, 5, 663-667, 2008.12, Non-alcoholic fatty liver disease (NAFLD) is considered to be associated with metabolic syndrome; however, a number of NAFLD patients are not obese. To explore any differences in lipid metabolism between obese and non-obese patients, we determined the expression of fatty acid metabolism-related genes. Expression levels of target genes were quantified by real-time PCR using liver biopsy samples from NAFLD patients and normal controls. Serum adipocytokine levels were also determined. The expression of genes related to fatty acid synthesis and uptake was generally up-regulated in NAFLD patients; however, no significant difference was seen between obese and non-obese groups. Most of the genes tested related to fatty acid and reactive oxygen species (ROS) elimination, were overexpressed in NAFLD and the levels were significantly higher in non-obese patients. As an exception, peroxisome proliferator-activated receptor a expression was suppressed in NAFLD and the levels were lower in the obese group. Triglyceride synthesis-related genes were up-regulated and lipolytic enzymes were decreased in NAFLD, but there was no significant difference between the obese and non-obese groups. In NAFLD, increased de novo synthesis and uptake of fatty acids led to further hepatocyte accumulation of fatty acids. The upregulation of fatty acid oxidation and the antioxidant pathway and the suppression of lipolysis seemed to be involved in this process. Expression of genes related to fatty acid oxidation and ROS elimination were higher in the non-obese group than in the obese group, which contributes to the trend of more severe liver injury, insulin resistance and steatosis in obese patients..
70. Nobito Higuchi, Masaki Kato, Yuki Shundo, Hirotaka Tajiri, Masatake Tanaka, Naoki Yamashita, Motoyuki Kohjima, Kazuhiro Kotoh, Makoto Nakamuta, Ryoichi Takayanagi, Munechika Enjoji, Liver X receptor in cooperation with SREBP-1c is a major lipid synthesis regulator in nonalcoholic fatty liver disease, Hepatology Research, 10.1111/j.1872-034X.2008.00382.x, 38, 11, 1122-1129, 2008.10, Aim: Nonalcoholic fatty liver disease (NAFLD) is one of the most frequent causes of liver dysfunction and its incidence has increased markedly. However, the mechanisms involved in the pathogenesis of NAFLD in humans have not been thoroughly investigated. Sterol regulatory element binding protein (SREBP)-1c and carbohydrate responsive element binding protein (ChREBP) are transcriptional factors that regulate the expression of lipogenic genes, including acetyl-CoA carboxylases (ACCs) and fatty acid synthase (FAS). SREBP-1c and ChREBP are transactivated by liver X receptor (LXR), a nuclear receptor that regulates the metabolism of cholesterol and fatty acids. To understand the mechanisms involved in the pathogenesis of NAFLD, we investigated the transcriptional factors and lipogenic genes activated in the liver with NAFLD. Methods: Real-time PCR was carried out on liver biopsy samples from 20 NAFLD patients. The target genes studied were: ACC1, FAS, SREBP-1c, ChREBP, AMP-activated protein kinase (AMPK), and LXRα. Results: LXRα, SREBP-1c, ACC1, and FAS were upregulated in NAFLD patients. Expression levels of LXR were four times greater than those of the controls and correlated significantly with SREBP-1c, but not with ChREBP, levels. Conclusions: These findings suggest that LXR acts as one of the main regulators of lipid metabolism by regulating SREBP-1c expression in NAFLD..
71. Munechika Enjoji, Kazuhiro Kotoh, Masaki Kato, Nobito Higuchi, Motoyuki Kohjima, Manabu Nakashima, Makoto Nakamuta, Therapeutic effect of ARBs on insulin resistance and liver injury in ptients with NAFLD and chronic hepatitis C
A pilot study, International journal of molecular medicine, 10.3892/ijmm_00000051, 22, 4, 521-527, 2008.10, Fatty liver is one of the local morphological manifestations of metabolic syndrome and is frequently associated with insulin resistance. Insulin resistance is also common in patients with chronic hepatitis C. Hyperinsulinemia is an independent risk factor for hypertension and cardiovascular mortality. The aim of this study was to evaluate the therapeutic efficacy of angiotensin II receptor blockers (ARBs), telmisartan and olmesartan, for patients with non-alcoholic fatty liver disease (NAFLD) and chronic hepatitis C (CH-C). We analyzed the incidence of obesity, insulin resistance, and other disorders in patients with NAFLD (Group A), CH-C (Group B), or other liver diseases (Group C). We evaluated whether the ARBs, telmisartan and olmesartan, improved insulin resistance and liver injury by measuring the homeostasis model assessment ratio of insulin resistance (HOMA-IR) and serum alanine aminotransferase (ALT). The incidence of obesity (BMI ≥25 kg/m2) was significantly higher in Group A than in Groups B and C. The incidence of insulin resistance (HOMA-IR ≥22.5) in Groups A and B was significantly higher than in Group C. Regular doses of telmisartan and olmesartan significantly improved HOMA-IR and ALT levels not only in NAFLD patients but also in patients with CH-C. The effects tended to be more notable with telmisartan. In conclusion, telmisartan and olmesartan improved insulin sensitivity and may possibly be used as liver protecting agents in CH-C as well as NAFLD patients..
72. Kazuhiro Kotoh, Munechika Enjoji, Masaki Kato, Motoyuki Kohjima, Makoto Nakamuta, Ryoichi Takayanagi, A new parameter using serum lactate dehydrogenase and alanine aminotransferase level is useful for predicting the prognosis of patients at an early stage of acute liver injury
A retrospective study, Comparative Hepatology, 10.1186/1476-5926-7-6, 7, 2008.08, Background: Although most patients with severe acute hepatitis are conservatively cured, some progress to acute liver failure (ALF) with a high rate of mortality. Based on the evidence that over-activation of macrophages, followed by disturbance of the hepatic microcirculation, plays a key role in ALF, we hypothesized that the production of serum lactate dehydrogenase (LDH) might increase in the liver under hypoxic conditions and could be an indicator to discriminate between conservative survivors and fatal patients at an early stage. Results: To confirm this hypothesis, we developed a new parameter with serum alanine aminotransferase (ALT) and LDH: the ALT-LDH index = serum ALT/(serum LDH - median of normal LDH range). We analyzed retrospectively 33 patients suffering acute liver injury (serum ALT more than 1000 U/L or prothrombin time expressed as international normalized ratio over 1.5 at admission) and evaluated the prognostic value of the ALT-LDH index, comparing data from the first 5 days of hospitalization with the Model for End-Stage Liver Disease (MELD) score. Patients whose symptoms had appeared more than 10 days before admission were excluded from this study. Among those included, 17 were conservative survivors, 9 underwent liver transplantation (LT) and 7 died waiting for LT. We found a rapid increase in the ALT-LDH index in conservative survivors but not in fatal patients. While the prognostic sensitivity and specificity of the ALT-LDH index was low on admission, at day 3 they were superior to the results of MELD. Conclusion: ALT-LDH index was useful to predict the prognosis of the patients with acute liver injury and should be helpful to begin preparation for LT soon after admission..
73. Motoyuki Kohjima, Nobito Higuchi, Masaki Kato, Kazuhiro Kotoh, Tsuyoshi Yoshimoto, Tatsuya Fujino, Masayoshi Yada, Ryoko Yada, Naohiko Harada, Munechika Enjoji, Ryoichi Takayanagi, Makoto Nakamuta, SREBP-1c, regulated by the insulin and AMPK signaling pathways, plays a role in nonalcoholic fatty liver disease, International journal of molecular medicine, 21, 4, 507-511, 2008.04, Nonalcoholic fatty liver disease (NAFLD) is a common liver disease whose prevalence has increased markedly. We reported previously that fatty acid synthesis was enhanced in NAFLD with the accumulation of fatty acids. To clarify the disorder, we evaluated the expression of genes regulating fatty acid synthesis by real-time PCR using samples from NAFLD (n=22) and normal liver (control; n=10). A major regulator of fatty acids synthesis is sterol regulatory element-binding protein-1c (SREBP-1c). Its expression was significantly higher in NAFLD, nearly 5-fold greater than the controls. SREBP-1c is positively regulated by insulin signaling pathways, including insulin receptor substrate (IRS)-1 and -2. In NAFLD, IRS-1 expression was enhanced and correlated positively with SREBP-1c expression. In contrast, IRS-2 expression decreased by 50% and was not correlated with SREBP-1c. Forkhead box protein A2 (Foxa2) is a positive regulator of fatty acid oxidation and is itself negatively regulated by IRSs. Foxa2 expression increased in NAFLD and showed a negative correlation with IRS-2, but not with IRS-1, expression. It is known that SREBP-1c is negatively regulated by AMP-activated protein kinase (AMPK) but expression levels of AMPK in NAFLD were almost equal to those of the controls. These data indicate that, in NAFLD, insulin signaling via IRS-1 causes the up-regulation of SREBP1-c, leading to the increased synthesis of fatty acids by the hepatocytes; negative feedback regulation via AMPK does not occur and the activation of Foxa2, following a decrease of IRS-2, up-regulates fatty acid oxidation..
74. Nobito Higuchi, Masaki Kato, Kazuhiro Kotoh, Motoyuki Kohjima, Shinichi Aishima, Makoto Nakamuta, Yoshinori Fukui, Ryoichi Takayanagi, Munechika Enjoji, Methylprednisolone injection via the portal vein suppresses inflammation in acute liver failure induced in rats by lipopolysaccharide and D-galactosamine, Liver International, 10.1111/j.1478-3231.2007.01590.x, 27, 10, 1342-1348, 2007.12, Background: We have reported that hepatic arterial steroid injection is an effective therapy to rescue patients from fulminant or severe acute hepatic failure. We speculate that a high concentration of steroid suppresses inflammatory processes in the liver directly by restraining activated inflammatory cells, including macrophages. To analyse the detailed mechanism, steroid injection via the portal vein was performed in an experimental model of liver damage. Methods: Rats subjected to lipopolysaccharide and D-galactosamine injection were treated with a methylprednisolone injection via the tail vein or the portal vein. The survival rate, serum levels of inflammatory cytokines and apoptotic cell counts in the liver were analysed. Results: The survival rate was significantly improved by steroid injection, especially via the portal vein. Serum values of alanine aminotransferase, tumor necrosis factor-α and interferon-γ were reduced in the treated groups, especially the group given portal venous injections. Apoptotic cell counts in the liver were significantly lower in the group injected with steroid via the portal vein. Conclusion: In the model rats, high concentrations of steroid in the liver acted on inflammatory cells and suppressed inflammatory cytokines and liver cell death. The mechanism is suggested to be the same for arterial steroid injection therapy in patients with acute hepatic failure..
75. Motoyuki Kohjima, Munechika Enjoji, Nobito Higuchi, Kazuhiro Kotoh, Masaki Kato, Ryoiichi Takayanagi, Makoto Nakamuta, NIM811, a nonimmunosuppressive cyclosporine analogue, suppresses collagen production and enhances collagenase activity in hepatic stellate cells, Liver International, 10.1111/j.1478-3231.2007.01560.x, 27, 9, 1273-1281, 2007.11, Background/Aims: A recent decrease in patient survival has been reported among hepatitis C virus (HCV)-infected liver transplant recipients and this may be attributable to progression of fibrosis. We reported previously that cyclosporine suppressed the proliferation of, and collagen production in, hepatic stellate cells (HSCs). Here, we investigated the effects of NIM811, a cyclosporine analogue, on cell growth, collagen production and collagenase activity in HSCs. Methods: Rat HSCs and human HSC-derived TWNT-4 cells were cultured for the study. The expression of collagen, matrix metalloproteinase 1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) and collagenase activity was evaluated. Cell proliferation and apoptosis were measured. Phosphorylation of mitogen-activated protein kinases (MAPKs), Smad2 and Smad3 was evaluated. The expression of the tumour growth factor-β (TGF-β)-receptor and Smad7 genes was also evaluated. Results: NIM811, as well as cyclosporine, suppressed the transcription and synthesis of collagen and stimulated the production of MMP-1 with a concomitant enhancement of collagenase activity, although it did not change the expression of TIMP-1. NIM811 inhibited proliferation without induction of apoptosis. In the MAPKs and TGF-β signalling pathways, NIM811 enhanced the phosphorylation of JNK and p38, but not extracellular signal-regulated kinases 1 and 2, and suppressed the phosphorylation of Smad2 and Smad3, accompanied by increased Smad7 transcription and decreased TGF-β-receptor transcription. Conclusion: These findings demonstrate that NIM811 not only suppresses collagen production and proliferation but also increases collagenase activity. These effects are accompanied by inhibition of TGF-β signalling pathways..
76. Motoyuki Kohjima, Munechika Enjoji, Nobito Higuchi, Masaki Kato, Kazuhiro Kotoh, Tsuyoshi Yoshimoto, Tatsuya Fujino, Masayoshi Yada, Ryoko Yada, Naohiko Harada, Ryoichi Takayanagi, Makoto Nakamuta, Re-evaluation of fatty acid metabolism-related gene expression in nonalcoholic fatty liver disease, International journal of molecular medicine, 20, 3, 351-358, 2007.09, Nonalcoholic fatty liver disease (NAFLD) is one of the most frequent causes of abnormal liver dysfunction, and its prevalence has markedly increased. We previously evaluated the expression of fatty acid metabolism-related genes in NAFLD and reported changes in expression that could contribute to increased fatty acid synthesis. In the present study, we evaluated the expression of additional fatty acid metabolism-related genes in larger groups of NAFLD (n=26) and normal liver (n=10) samples. The target genes for real-time PCR analysis were as follows: acetyl-CoA carboxylase (ACC) 1, ACC2, fatty acid synthase (FAS), sterol regulatory element-binding protein 1c (SREBP-1c), and adipose differentiation-related protein (ADRP) for evaluation of de novo synthesis and uptake of fatty acids; carnitine palmitoyltransferase 1a (CPT1a), long-chain acyl-CoA dehydrogenase (LCAD), long-chain L-3-hydroxyacylcoenzyme A dehydrogenase α (HADHα), uncoupling protein 2 (UCP2), straight-chain acyl-CoA oxidase (ACOX), branched-chain acyl-CoA oxidase (BOX), cytochrome P450 2E1 (CYP2E1), CYP4A11, and peroxisome proliferator-activated receptor (PPAR)a for oxidation in the mitochondria, peroxisomes and microsomes; superoxide dismutase (SOD), catalase, and glutathione synthetase (GSS) for antioxidant pathways; and diacylglycerol O-acyltransferase 1 (DGAT1), PPARγ, and hormone-sensitive lipase (HSL) for triglyceride synthesis and catalysis. In NAFLD, although fatty acids accumulated in hepatocytes, their de novo synthesis and uptake were up-regulated in association with increased expression of ACC1, FAS, SREBP-1c, and ADRP. Fatty acid oxidation-related genes, LCAD, HADHα, UCP2, ACOX, BOX, CYP2E1, and CYP4A11, were all overexpressed, indicating that oxidation was enhanced in NAFLD, whereas the expression of CTP1a and PPARα was decreased. Furthermore, SOD and catalase were also overexpressed, indicating that antioxidant pathways are activated to neutralize reactive oxygen species (ROS), which are overproduced during oxidative processes. The expression of DGAT1 was up-regulated without increased PPARγ expression, whereas the expression of HSL was decreased. Our data indicated the following regarding NAFLD: i) increased de novo synthesis and uptake of fatty acids lead to further fatty acid accumulation in hepatocytes; ii) mitochondrial fatty acid oxidation is decreased or fully activated; iii) in order to complement the function of mitochondria (β-oxidation), peroxisomal (β-oxidation) and microsomal (ω-oxidation) oxidation is up-regulated to decrease fatty acid accumulation; iv) antioxidant pathways including SOD and catalase are enhanced to neutralize ROS overproduced during mitochondrial, peroxisomal, and microsomal oxidation; and v) lipid droplet formation is enhanced due to increased DGAT expression and decreased HSL expression. Further studies will be needed to clarify how fatty acid synthesis is increased by SREBP-1c, which is under the control of insulin and AMP-activated protein kinase..
77. Sawako Inoue, Shusuke Morizono, Shinsaku Yamashita, Yuki Horikawa, Motoyuki Kohjima, Yuzuru Miyagi, Tsuyosi Yoshimoto, Kazuhiro Kotoh, Munechika Enjoji, Ryoichi Takayanagi, Yuji Soejima, Akinobu Taketomi, Tomoharu Yoshizumi, Yoshihiko Maehara, Makoto Nakamuta, Estimation of indication for living donor liver transplantation in patients with HCV and/or HBV infection, Fukuoka igaku zasshi = Hukuoka acta medica, 98, 7, 295-300, 2007.07, We evaluated 78 patients with chronic viral hepatitis for liver transplantation. 51 patients met our original criteria for liver transplantation, and 35 patients of them suffered from hepatocellular carcinoma (HCC). Patients with HCC were significantly older and showed higher prothrombin activity than those without HCC. Eighteen of 35 patients with HCC did not meet the Milan criteria, and they showed lower levels of total bilirubin, Child-Pugh score, and MELD score than those who met the criteria. Theses results indicate that acceptability for transplantation should be evaluated soon after the patients have become candidates for liver transplantation. In Japan, decompensated liver cirrhosis is a necessary condition for the application of public health insurance against liver transplantarion and, in cases with HCC, it is necessary to meet the Milan criteria. Application to liver transplantation should also be considered based on HCC stage such as the UNOS scoring system..
78. Tsuyoshi Yoshimoto, Kazuhiro Kotoh, Yuki Horikawa, Motoyuki Kohjima, Shusuke Morizono, Shinsaku Yamashita, Munechika Enjoji, Makoto Nakamuta, Decreased portal flow volume increases the area of necrosis caused by radio frequency ablation in pigs, Liver International, 10.1111/j.1478-3231.2007.01454.x, 27, 3, 368-372, 2007.04, Background/aims: Although radio frequency ablation (RFA) has been widely accepted as an effective treatment for hepatocellular carcinoma (HCC), severe complications are not uncommon. Major complications seem to occur as a result of over-ablation beyond the intended area. As most patients with HCC have underlying cirrhosis, we speculated that decreased portal flow might cause the necrosis associated with RFA. To confirm this hypothesis, we examined the area of necrosis resulting from RFA under varying conditions of portal flow in a porcine model. Methods: RFA was performed using ultrasonographic guidance in anesthetized pigs. During the RFA procedure, portal flow was regulated by a balloon catheter, which was set in a portal trunk. The necrosis area was measured after sacrifice and was compared with the hyperechoic area that appeared during ablation. In another session, RFA was performed close to the hepatic vein and endothelial damage was examined. Results: The necrosis area caused by RFA was significantly larger when the portal flow volume was decreased by 50% or more. The hyperechoic lesion was always larger than the area of pathological necrosis regardless of portal flow volume. Under conditions of decreased portal flow, the vessel endothelium near the ablated area was more readily damaged. Conclusion: Decreased portal flow volume resulted in enlargement of the area of necrosis caused by RFA. Our results indicate that over-ablation could easily occur in patients with advanced cirrhosis, and that this could lead to major complications. Ultrasonographic guidance may be helpful for avoiding over-ablation..
79. Makoto Nakamuta, Motoyuki Kohjima, Shusuke Morizono, Tsuyoshi Yoshimoto, Yuzuru Miyagi, Hironori Sakai, Munechika Enjoji, Kazuhiro Kotoh, Comparison of tissue pressure and ablation time between the LeVeen and cool-tip needle methods, Comparative Hepatology, 10.1186/1476-5926-5-10, 5, 2006.12, Background: Radio frequency ablation (RFA) has been accepted clinically as a useful local treatment for hepatocellular carcinoma (HCC). However, intrahepatic recurrence after RFA has been reported which might be attributable to increase in intra-tumor pressure during RFA. To reduce the pressure and ablation time, we developed a novel method of RFA, a multi-step method in which a LeVeen needle, an expansion-type electrode, is incrementally and stepwise expanded. We compared the maximal pressure during ablation and the total ablation time among the multi-step method, single-step method (a standard single-step full expansion with a LeVeen needle), and the method with a cool-tip electrode. Finally, we performed a preliminary comparison of the ablation times for these methods in HCC cases. Results: A block of pig liver sealed in a rigid plastic case was used as a model of an HCC tumor with a capsule. The multi-step method with the LeVeen electrode resulted in the lowest pressure as compared with the single-step or cool-tip methods. There was no significant difference in the ablation time between the multi-step and cool-tip ablation methods, although the single-step methods had longer ablation times than the other ablation procedures. In HCC cases, the multi-step method had a significantly shorter ablation time than the single-step or cool-tip methods. Conclusion: We demonstrated that the multi-step method was useful to reduce the ablation time and to suppress the increase in pressure. The multi-step method using a LeVeen needle may be a clinically applicable procedure for RFA..
80. Tsuyoshi Yoshimoto, Makoto Nakamuta, Kazuhiro Kotoh, Motoyuki Kohjima, Shusuke Morizono, Yuzuru Miyagi, Hironori Sakai, Munechika Enjoji, An adult case with hunter's syndrome presenting prominent hepatic failure
Light and electron microscopic features of the liver, Internal Medicine, 10.2169/internalmedicine.45.6005, 45, 20, 1133-1135, 2006.11, We describe a 40-year-old male patient with Hunter's syndrome. His main manifestations were ascites and esophageal varices due to cirrhotic liver. We obtained hepatic biopsy samples and examined them. Ultrastructurally, the features of the hepatocytes and Kupffer cells were the same as those reported in young patients. The passage of 40 years led to gradual progression to fibrosis, and ultimately liver cirrhosis. Namely, with a longer survival time, the complications of liver cirrhosis become more remarkable. Hepatic fibrosis in Hunter's syndrome is slowly progressive and patients who are expected to have a longer life span should be continuously monitored for hepatic complications..
81. Kazuhiro Kotoh, Munechika Enjoji, Makoto Nakamuta, Tsuyoshi Yoshimoto, Motoyuki Kohjima, Shusuke Morizono, Shinsaku Yamashita, Yuki Horikawa, Kengo Yoshimitsu, Tsuyoshi Tajima, Yoshiki Asayama, Kousei Ishigami, Masakazu Hirakawa, Arterial steroid injection therapy can inhibit the progression of severe acute hepatic failure toward fulminant liver failure, World Journal of Gastroenterology, 10.3748/wjg.v12.i41.6678, 12, 41, 6678-6682, 2006.11, Aim: To utilize transcatheter arterial steroid injection theraphy (TASIT) via the hepatic artery to reduce hepatic macrophage activity in patients with severe acute hepatic failure. Methods: Thirty-four patients with severe acute hepatic failure were admitted to our hospital between June 2002 to June 2006 providing for the possibility of liver transplantation (LT). Seventeen patients were treated using traditional liver supportive procedures, and the other 17 patients additionally underwent TASIT with 1000 mg methylprednisolone per day for 3 continuous days. Results: Of the 17 patients who received TASIT, 13 were cured without any complications, 2 died, and 2 underwent LT. Of the 17 patients who did not receive TASIT, 4 were self-limiting, 7 died, and 6 underwent LT. Univariate logistic analysis revealed that ascites, serum albumin, prothrombin time, platelet count, and TASIT were significant variables for predicating the prognosis. Multivariate logistic regression analysis using stepwise variable selection showed that prothrombin time, platelet count, and TASIT were independent predictive factors. Conclusion: TASIT might effectively prevent the progression of severe acute hepatic failure to a fatal stage of fulminant liver failure..
82. Munechika Enjoji, Kazuhiro Kotoh, Manabu Nakashima, Tsuyoshi Yoshimoto, Yuzuru Miyagi, Motoyuki Kohjima, Makoto Nakamuta, RCAS1-expressing macrophages in inflammatory liver diseases [3], Liver International, 10.1111/j.1478-3231.2005.01223.x, 26, 3, 385-387, 2006.04.
83. Tomoko Izaki, Sachiko Kamakura, Motoyuki Kohjima, Hideki Sumimoto, Two forms of human Inscuteable-related protein that links Par3 to the Pins homologues LGN and AGS3, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2006.01.050, 341, 4, 1001-1006, 2006.03, In cell polarization of Drosophila neuroblasts, Inscuteable (Insc) functions via tethering Partner of Insc (Pins) to Bazooka, homologous to human cell polarity protein Par3. However, little has been known about mammalian homologues of Insc. Here we describe cloning of two distinct cDNAs from human Insc gene, which is differentially expressed from alternative first exons: one encodes 579 amino acids, whereas the other lacks the N-terminal 47 amino acids. In contrast to human homologues for Pins and Par3, human Insc exhibits a weak homology with the Drosophila counterpart. Nevertheless, human Insc proteins bind to the human Pins homologues LGN and AGS3, and also to human Par3 and its related protein Par3β. Although LGN by itself is incapable of interacting with Par3, coexpression of human Insc leads to the interaction between LGN and Par3, indicating that human Insc plays an evolutionarily conserved role as an adaptor protein that links Pins to Par3..
84. Makoto Nakamuta, Shusuke Morizono, Motoyuki Kohjima, Kazuhiro Kotoh, Munechika Enjoji, Baseline characterization of patients aged 70 years and above with hepatocellular carcinoma, World Journal of Gastroenterology, 10.3748/wjg.v11.i47.7512, 11, 47, 7512-7514, 2005.12, Aim: To characterize the baseline profiles of patients aged 70 years and above with hepatocellular carcinoma (HCC). Methods: A series of 127 consecutive patients with HCC were enrolled between 2000 and 2004, and none of them had been diagnosed as having HCC previously. Baseline profiles, including parameters of hepatic function such as serum transaminase and prothrombin time [PT (% activity)] were compared between patients aged ≥70 and <70 years. Results: Patients ≥70 years old showed significantly lower levels of aspartate aminotransferase (P = 0.04) and alanine aminotransferase (P = 0.01), and significantly higher PTs (P = 0.04) and platelet counts (P = 0.02). Concomitantly, among ≥70-year-old patients, HCC was more common in non-cirrhotics, whereas among patients <70 years old, HCC was more common in cirrhotics. There was no significant difference between the groups in the number or size of tumors. Conclusion: Older HCC patients showed less inflammation and better preservation of hepatic function, indicating that not only cirrhotic patients but also non-cirrhotic patients should be considered as a high-risk group among the elderly..
85. M. Nakamuta, M. Kohjima, M. Fukushima, S. Morizono, K. Kotoh, N. Kobayashi, M. Enjoji, Cyclosporine suppresses cell growth and collagen production in hepatic stellate cells, Transplantation Proceedings, 10.1016/j.transproceed.2005.10.104, 37, 10, 4598-4602, 2005.12, Background. In HCV-related graft hepatitis, immunosuppression has been implicated in rapid progression to cirrhosis, a serious clinical issue. We investigated the effects of cyclosporine or tacrolimus on cell growth and collagen production by hepatic stellate cells (HSC), which play a role in hepatic fibrosis. Materials and methods. Cultured rat HSCs and human HSC-derived TWNT-4 cells were evaluated for proliferation, type I collagen, phosphorylation states of mitogen-activated protein kinases extracellular signal-regulated kinase 1/2; [MAPKs Erk1/2], c-Jun N-terminal kinase (JNK, p38), as well as the expression of collagen, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) genes. Results. Cyclosporine suppressed cell growth and collagen production in a concentration-dependent manner. At clinically relevant concentrations of 0.125 μmol (150 ng/mL) cyclosporine significantly reduced collagen production per cell by more than 50%. Similarly, tacrolimus also reduced both collagen concentration and cell number; however, tacrolimus at a clinically relevant concentration of 12.5 nmol (10 ng/mL) did not significantly reduce collagen production. Treatment with cyclosporine reduced type I collagen and TIMP-1 expression and enhanced MMP-1 expression. Cyclosporine also inhibited phosphorylation strongly for JNK and p38, and weakly inhibited for Erk1/2. Conclusion. These findings demonstrated that cyclosporine suppresses cell growth and collagen production, suggesting that it may have an antifibrogenic effect..
86. Kazuhiro Kotoh, Shusuke Morizono, Motoyuki Kohjima, Munechika Enjoji, Hironori Sakai, Makoto Nakamuta, Evaluation of liver parenchymal pressure and portal endothelium damage during radio frequency ablation in an in vivo porcine model, Liver International, 10.1111/j.1478-3231.2005.01167.x, 25, 6, 1217-1223, 2005.12, Background/aims: We previously developed a multi-step, incremental expansion method (multi-step method) for radio frequency ablation (RFA) in vitro, which prevented increases in pressure and reduced the ablation time as compared with other methods. In this study, we evaluated liver parenchymal pressure and portal endothelium damage during RFA with different devices and protocols in an in vivo porcine model. Method: Nine healthy female pigs were anaesthetized. RFA was performed with two different devices and protocols; one involved the use of a LeVeen needle with a single-step full expansion method or a multi-step method, and the other used a cool-tip needle with 40 or 60 W power. We measured the pressure in the liver parenchyma and the gallbladder during RFA. We also evaluated portal endothelium damage by NADH staining. Results: The multi-step method with the LeVeen electrode resulted in the lowest parenchymal and intra-gallbladder pressures (multi-step method < single-step method < cool-tip 40 W < cool-tip 60 W). In contrast, the ablation time was shortest with the cool-tip needle at 60 W (cool-tip 60 W
87. Marie Fukushima, Munechika Enjoji, Motoyuki Kohjima, Rie Sugimoto, Satoshi Ohta, Kazuhiro Kotoh, Masami Kuniyoshi, Kunihisa Kobayashi, Minako Imamura, Toyoshi Inoguchi, Makoto Nakamuta, Hajime Nawata, Adipose differentiation related protein induces lipid accumulation and lipid droplet formation in hepatic stellate cells, In Vitro Cellular and Developmental Biology - Animal, 10.1290/0410069.1, 41, 10, 321-324, 2005.11, The function of adipose differentiation-related protein (ADRP) is known to be the uptake of long-chain fatty acids and formation of lipid droplets in lipid-accumulating cells. We hypothesized that ADRP might stimulate activated hepatic stellate cells (HSCs) to accumulate lipids, resulting in their transition to the quiescent state. In this study, cultured HSCs in fifth passages isolated from rat were infected by adenovirus vector expressing ADRP (Ad.GFP-ADRP), and morphologic and functional changes were evaluated in comparison with control HSCs infected by recombinant adenovirus-expressing β-galactosidase (Ad.LacZ). In Ad.GFP-ADRP-infected cells only, many tiny lipid droplets were apparent in the cytoplasm, while the outline of the cells was not changed. The ADRP was detected around the lipid droplets. In HSCs with intracellular actin filaments, the staining pattern of the filaments before and after infection with Ad.GFP-ADRP or Ad.LacZ did not differ. The cell proliferation rate was not influenced by infection with Ad.LacZ or Ad.GFP-ADRP. Type I collagen secretion from cells overexpressing ADRP was not significantly different from that of Ad.LacZ-infected cells. In our in vitro study, ADRP overexpression induced the formation of cytoplasmic lipid droplets in activated HSCs but could not convert other characteristics of the activated form into those of the quiescent form..
88. Kazuhiro Kotoh, Munechika Enjoji, Eiichirou Arimura, Shusuke Morizono, Motoyuki Kohjima, Hironori Sakai, Makoto Nakamuta, Scattered and rapid intrahepatic recurrences after radio frequency ablation for hepatocellular carcinoma, World Journal of Gastroenterology, 10.3748/wjg.v11.i43.6828, 11, 43, 6828-6832, 2005.11, Aim: To evaluate a series of patients with hepatocellular carcinoma (HCC) treated with several different protocols and devices. Methods: We treated 138 patients [chronic hepatitis/liver cirrhosis (Child-Pugh A/B/C), 3/135 (107/25/3)] with two different devices and protocols: cool-tip needle [initial ablation at 60 W (standard method) (n = 37) or at 40 W (modified method) (n = 28)] or; ablation with a LeVeen needle using a standard single-step, full expansion (single-step) method (n = 39) or a multi-step, incremental expansion (multi-step) method. Results: Eleven patients experienced rapid and scattered recurrences 1 to 7 mo after the ablation. Nine patients were treated by the cool-tip original protocol (60 W) (9/37 = 24%) and the other two by the LeVeen single-step method (2/39 = 5%). The location of the recurrence was surrounding and limited to the site of ablation segment in three cases, and spread over one lobule or both lobules in the other eight cases. There was no recurrence in the patients treated with the modified cool-tip modified method (40 W) or the LeVeen multi-step method. Conclusion: There is a risk of rapid and scattered recurrence after RFA, especially when the standard cool-tip procedure is used. Because such recurrence would worsen the prognosis, we recommend that modified protocols for the cool-tip and LeVeen needle methods should be used in clinical practice..
89. Satoshi Ohta, Makoto Nakamuta, Marie Fukushima, Motoyuki Kohjima, Kazuhiro Kotoh, Munechika Enjoji, Hajime Nawata, Beraprost sodium, a prostacyclin (PGI2) analogue, ameliorates concanavalin A-induced liver injury in mice, Liver International, 10.1111/j.1478-3231.2005.01143.x, 25, 5, 1061-1068, 2005.10, Background/Aims: Prostacyclin (PGI2) is a potent mediator in the inflammatory and coagulation processes. The aim of this study was to test whether beraprost sodium, a PGI2 analogue, could prevent experimental hepatic injury induced by concanavalin A (Con A), which is a model of fulminant hepatic failure. Methods: Beraprost (100 μg/kg) was administered intraperitoneally simultaneously with Con A (40 mg/kg) in C57B6J mice. Blood circulation in the liver was determined by laser-Doppler flowmetry. Plasma levels of alanine aminotransferase (ALT), tumor necrosis factor (TNF)-α, interferon (IFN)-γ, and interleukin (IL)-6 were determined. Levels of TNF-α and IFN-γ in culture supernatant of splenocytes were also determined. Results: Beraprost administration reduced the incidence of death following hepatic failure (76.5% vs. 29.4%, P<0.05). Plasma levels of ALT were significantly lower in the beraprost-treated group than in the control group, and in the former, there was concomitant suppression of the histological features of injury. Beraprost significantly increased hepatic blood flow volume in Con A-treated mice. Plasma levels of TNF-α and IFN-γ were significantly reduced at 6 and 12h after Con A injection, respectively, but the levels of IL-6 were increased at 6h. In vitro, beraprost also suppressed Con A-induced TNF-α production in splenocytes, while it stimulated IFN-γ production. Conclusion: These findings imply that beraprost suppresses Con A-induced liver injury. These data also suggest that beraparost, which is clinically effective in treating pulmonary hypertension, may have therapeutic potential for preventing hepatic injury..
90. Makoto Nakamuta, Nobuhiko Higashi, Motoyuki Kohjima, Marie Fukushima, Satoshi Ohta, Kazuhiro Kotoh, Naoya Kobayashi, Munechika Enjoji, Epigallocatechin-3-gallate, a polyphenol component of green tea, suppresses both collagen production and collagenase activity in hepatic stellate cells., International journal of molecular medicine, 16, 4, 677-681, 2005.10, Catechins such as epigallocatechin-3-gallate (EGCG), epicatechin-3-gallate (ECG), and epigallocatechin (EGC) are polyphenol components of green tea. EGCG is the major component and has been reported to possess a wide range of biological properties including anti-fibrogenic activity. In hepatic fibrosis, activated hepatic stellate cells (HSCs) play a central role. In this study, we investigated the effect of catechins, including EGCG, on collagen production and collagenase activity in rat primary HSCs and activated human HSC-derived TWNT-4 cells. EGCG (50 microM) suppressed type I collagen production in rat HSCs more than ECG (50 microM) did; however, EGC (50 microM) did not show suppressive effects. EGCG also inhibited both collagen production and collagenase activity (active matrix metalloproteinase-1 [MMP-1]) in a dose-dependent manner, but did not affect the tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) production in TWNT-4 cells. Real-time PCR unexpectedly revealed that EGCG enhanced the transcription of type I collagen and TIMP-1, but did not affect the transcription of alpha-smooth muscle actin (alpha-SMA), and reduced the transcription MMP-1 in TWNT-4 cells. These findings demonstrated that EGCG inhibited collagen production regardless of enhanced collagen transcription and suppressed collagenase activity, and suggested that EGCG might have therapeutic potential for liver fibrosis..
91. Makoto Nakamuta, Motoyuki Kohjima, Shusuke Morizono, Kazuhiro Kotoh, Tsuyoshi Yoshimoto, Izuru Miyagi, Munechika Enjoji, Evaluation of fatty acid metabolism-related gene expression in nonalcoholic fatty liver disease., International journal of molecular medicine, 16, 4, 631-635, 2005.10, Nonalcoholic fatty liver disease (NAFLD) is one of the most frequent causes of abnormal liver dysfunction, and its prevalence has markedly increased; however, the mechanisms involved in the pathogenesis of NAFLD have not been thoroughly investigated in humans. In this study, we evaluated the expression of fatty acid metabolism-related genes in NAFLD. Real-time RT-PCR was performed using liver biopsy samples from 12 NAFLD patients. The target genes studied were: acetyl-CoA carboxylase (ACC) 1, ACC2, and fatty acid synthase (FAS) for the evaluation of de novo fatty acid synthesis; carnitine palmitoyltransferase 1a (CPT1a), long-chain acyl-CoA dehydrogenase (LCAD), and long-chain L-3-hydroxyacyl-coenzyme A dehydrogenase alpha (HADHalpha) for beta-oxidation in the mitochondria; peroxisome proliferator-activated receptor- (PPAR-) alpha and cytochrome P450 2E1 (CYP2E1) for oxidation in peroxisomes and microsomes (endoplasmic reticulum) respectively; and diacylglycerol O-acyltransferase 1 (DGAT1), PPAR-gamma, and hormone sensitive lipase (HSL) for triglyceride synthesis and catalysis. In NAFLD, expression of ACC1 and ACC2, but not FAS was increased, indicating that de novo fatty acid synthesis is enhanced in NAFLD. In contrast, expression of CTP1a, a rate-limiting enzyme, was remarkably decreased, indicating that beta-oxidation in the mitochondria was decreased, although the expression of LCAD and HADHalpha was increased. Expression of PPAR-alpha was increased, whereas that of CYP2E1 was reduced. The expression of DGAT1, PPAR-gamma, and HSL was enhanced. These data suggest that in NAFLD, increased de novo synthesis and decreased beta-oxidation in the mitochondria lead to accumulation of fatty acids in hepatocytes, although the extent of oxidation in peroxisomes and microsomes remains unclear..
92. Rie Sugimoto, Munechika Enjoji, Motoyuki Kohjima, Satoshi Tsuruta, Marie Fukushima, Masataka Iwao, Toshiyo Sonta, Kazuhiro Kotoh, Toyoshi Inoguchi, Makoto Nakamuta, High glucose stimulates hepatic stellate cells to proliferate and to produce collagen through free radical production and activation of mitogen-activated protein kinase, Liver International, 10.1111/j.1478-3231.2005.01130.x, 25, 5, 1018-1026, 2005.10, Background: Nonalcoholic steatohepatitis is a clinicopathologic condition that may progress to liver fibrosis. Hyperglycemia is supposed to be one of the factors inducing hepatic fibrogenesis, but the mechanism has not been fully clarified. Oxidative stress is increasingly found in patients with diabetes/hyperglycemia in which conditions reactive oxygen species (ROS) are produced. Methods: We performed experiments using hepatic stellate cells (HSCs) in culture in order to confirm the effect of high glucose concentrations on cell proliferation, type I collagen production, ROS production and activation of mitogen-activated protein (MAP) kinase pathway. Results: High glucose stimulated cell growth of HSCs and upregulated the levels of activated/phosphorylated extracellular signal-regulated kinase 1/2 and free radical production in HSCs. The MAP kinase phosphorylation and cell proliferation were suppressed by diphenylene iodonium chloride, an NADPH oxidase inhibitor, and by calphostin C, a protein kinase C (PKC)-specific inhibitor. Increased type I collagen mRNA and protein levels were also observed in HSCs at high glucose concentrations. Conclusions: Our findings indicate that high glucose concentrations may stimulate ROS production through PKC-dependent activation of NADPH oxidase, and induce MAP kinase phosphorylation subsequent to proliferation and type I collagen production by HSCs..
93. Munechika Enjoji, Shusuke Morizono, Kazuhiro Kotoh, Motoyuki Kohjima, Yuzuru Miyagi, Tsuyoshi Yoshimoto, Makoto Nakamuta, Re-evaluation of antitumor effects of combination chemotherapy with interferon-α and 5-fluorouracil for advanced hepatocellular carcinoma, World Journal of Gastroenterology, 10.3748/wjg.v11.i36.5685, 11, 36, 5685-5687, 2005.09, Aim: To evaluate the efficacy of combination chemotherapy with interferon-α (IFNα) and 5-fluorouracil (5-FU) in patients with advanced hepatocellular carcinoma (HCC). Methods: Twenty-eight HCC patients in advanced stage were enrolled in the study. They were treated with IFNα/5-FU combination chemotherapy. One cycle of therapy lasted for 4 wk. IFNα (3×106 units) was subcutaneously injected thrice weekly on days 1, 3, and 5 for 3 wk, and 5-FU (500 mg/d) was administered via the proper hepatic artery for 5 consecutive days per week for 3 wk. No drugs were administered during the 4th wk. The effect of combination chemotherapy was evaluated in each patient after every cycle based on the reduction of tumor volume. Results: After the 1st cycle of therapy, 16 patients showed a partial response (PR, 57.1%) but none showed a complete response (CR, 0%). At the end of therapy, the number of patients who showed a CR, PR, or no response (NR) was 1, 10, and 17, respectively. The response rate for therapy (CR+PR) was 21.5%. Biochemical tests before therapy were compared between responsive (CR+PR) and non-responsive (NR) patients, but no significant differences were found for any of the parameters examined, indicating that no reasonable predictors could be identified in our analysis. Conclusion: Attempts should be made to discriminate between responders and non-responders by evaluating tumor size after the first cycle of IFNα/5-FU combination chemotherapy. For non-responders, therapy should not proceed to the next cycle, and instead, different combination of anticancer drugs should be explored..
94. Marie Fukushima, Makoto Nakamuta, Motoyuki Kohjima, Kazuhiro Kotoh, Munechika Enjoji, Naoya Kobayashi, Hajime Nawata, Fasudil hydrochloride hydrate, a Rho-kinase (ROCK) inhibitor, suppresses collagen production and enhances collagenase activity in hepatic stellate cells, Liver International, 10.1111/j.1478-3231.2005.01142.x, 25, 4, 829-838, 2005.08, Background/Aims: The Rho-ROCK signaling pathways play an important role in the activation of hepatic stellate cells (HSCs). We investigated the effects of fasudil hydrochloride hydrate (fasudil), a Rho-kinase (ROCK) inhibitor, on cell growth, collagen production, and collagenase activity in HSCs. Methods: Rat HSCs and human HSC-derived TWNT-4 cells were cultured for studies on stress fiber formation and α-smooth muscle actin (α-SMA) expression. Proliferation was measured by BrdU incorporation, and apoptosis by TUNEL assay. The phosphorylation states of the MAP kinases (MAPKs), extra cellular signal-regulated kinase 1/2 (ERK1/2), c-jun kinase (JNK), and p38 were evaluated by western blot analysis. Type I collagen, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) production and gene expression were evaluated by ELISA and real-time PCR, respectively. Collagenase activity (active MMP-1) was also evaluated. Results: Fasudil (100 μM) inhibited cell spreading, the formation of stress fibers, and expression of α-SMA with concomitant suppression of cell growth, although it did not induce apoptosis. Fasudil inhibited phosphorylation of ERK1/2, JNK, and p38. Treatment with fasudil suppressed the production and transcription of collagen and TIMP, stimulated the production and transcription of MMP-1, and enhanced collagenase activity. Conclusion: These findings demonstrated that fasudil not only suppresses proliferation and collagen production but also increases collagenase activity..
95. Makoto Nakamuta, Shusuke Morizono, Satoru Tsuruta, Motoyuki Kohjima, Kazuhiro Kotoh, Munechika Enjoji, Remote delivery and expression of soluble type II TGF-beta receptor in muscle prevents hepatic fibrosis in rats., International journal of molecular medicine, 16, 1, 59-64, 2005.07, Transforming growth factor-beta (TGF-beta) has been implicated in the process of hepatic fibrosis, and stimulates production of extracellular matrix in hepatic stellate cells, which play a major role in the process. It has been recently reported that blockage of TGF-beta signaling prevents hepatic fibrosis. We evaluated a strategy for anti-TGF-beta gene therapy for hepatic fibrosis by transfecting plasmids expressing an entire extracellular domain of human TGF-beta type II [soluble type II TGF-beta receptor (sTGF-betaIIR)] into skeletal muscle in a rat experimental model of dimethylnitrosamine- (DMN-) induced fibrosis. sTGF-betaIIR treatment decreased significantly the occurrence of DMN-induced hepatic fibrosis, evaluated by computed image analysis and by measurement of hydroxyproline content of the liver, and reduced the expression of collagen and alpha-smooth muscle actin. The treatment also caused a significant decrease in hepatic levels of interleukin- (IL-) 12 (Th1 cytokine) and an increase in those of IL-10 (Th2 cytokine), indicating a change in the Th1/Th2 cytokine balance in the liver. In conclusion, blockade of TGF-beta after intramuscular transfer of the soluble type II TGF-beta receptor gene suppressed hepatic fibrosis, suggesting that this strategy may be useful for gene therapy of hepatic fibrosis..
96. Eiichirou Arimura, Makoto Nakamuta, Kazuhiro Kotoh, Shusuke Morizono, Motoyuki Kohjima, Marie Fukushima, Munechika Enjoji, Hajime Nawata, A case of idiopathic portal hypertension complicated with sepsis caused by Aeromonas hydrophila, Fukuoka igaku zasshi = Hukuoka acta medica, 96, 6, 259-264, 2005.06, A 53 year old man with idiopathic portal hypertension (IPH) was admitted because of high fever and diarrhea. Nineteen years before admission, he had received splenic hilar renal shunt operation with proximal flush ligation of splenic vein due to gastric varices. Three months before admission, he had been admitted to our hospital for evaluation of liver dysfunction. Liver biopsy examination had revealed peri-portal fibrosis consistent with IPH. Aeromonas hydrophila was isolated from blood. Although he was treated with antibiotics plus dopamine, glucose-insulin therapy, and mechanical ventilation, he had severe septic shock, and died 29 days after admission. We have to take notice of A. hydrophila infection in cases of portosystemic shunt because they fall in severe septic shock..
97. Kazuhiro Kotoh, Makoto Nakamuta, Shusuke Morizono, Motoyuki Kohjima, Eiichirou Arimura, Marie Fukushima, Munechika Enjoji, Hironori Sakai, Hajime Nawata, A multi-step, incremental expansion method for radio frequency ablation
Optimization of the procedure to prevent increases in intra-tumor pressure and to reduce the ablation time, Liver International, 10.1111/j.1478-3231.2005.01051.x, 25, 3, 542-547, 2005.06, Background/Aims: Radio frequency ablation (RFA) has been accepted clinically as a useful local treatment for hepatocellular carcinoma (HCC). However, intra-hepatic recurrence after RFA has been reported. We initially hypothesized that recurrence was attributable to increases in intratumor pressure during RFA, and we subsequently measured the pressure and optimized the procedure. Methods: A block of pig liver sealed in a rigid plastic case was used as a model of an HCC tumor with a capsule. We compared the pressure between a single-step full expansion of the needle (single-step method) and incremental, stepwise expansion (multi-step method), and evaluated the effect of varying the electrical power. Finally, we performed a preliminary comparison of the ablation times for these methods in HCC cases. Results: The multi-step method resulted in a significantly lower pressure and shorter total ablation time than the single-step method. Furthermore, incremental expansion in 10 steps resulted in a lower pressure and shorter ablation time than four steps. Seventy W-ablation resulted in a lower pressure and shorter time than 30- or 50 W-ablation. In HCC cases, the multiple-step method had a significantly shorter ablation time than the single-step method. Conclusion: The multi-step method can be recommended to reduce the ablation time, and suppress the increase in pressure..
98. Nobuhiko Higashi, Motoyuki Kohjima, Marie Fukushima, Satoshi Ohta, Kazuhiro Kotoh, Munechika Enjoji, Naoya Kobayashi, Makoto Nakamuta, Epigallocatechin-3-gallate, a green-tea polyphenol, suppresses Rho signaling in TWNT-4 human hepatic stellate cells, Journal of Laboratory and Clinical Medicine, 10.1016/j.lab.2005.03.017, 145, 6, 316-322, 2005.06, Epigallocatechin-3-gallate (EGCG), a major constituent of the polyphenoids in green tea, has been reported to possess a wide range of biologic activities, including antifibrogenesis. Activated hepatic stellate cells (HSCs) are central to hepatic fibrosis, and Rho (a small GTPase)-signaling pathways have been implicated in the activation and proliferation of HSCs. In this study, we investigated the effect of EGCG on Rho-signaling pathways in activated human HSC-derived TWNT-4 cells. EGCG inhibited stress-fiber formation, an indicator of Rho activation, and changed the distribution of α-smooth-muscle actin. These inhibitory effects of EGCG were restored by overexpression of constitutively active Rho. A pull-down assay revealed that activated Rho (GTP-bound state) was strongly inhibited by ECGC and accompanied by suppressed phosphorylation of focal adhesion kinase, which is a regulator of Rho-signaling pathways. 5-Bromo-2′-deoxy-uridine incorporation demonstrated that ECGC (100 μmol/L suppressed cell growth by 80%, and terminal deoxynucleotidyl transferase viotin-deoxyruidine triphosphate nick end-labeling revealed that EGCG (100 μmol/L) caused apoptosis in half of the total cells. EGCG also strongly inhibited lysophoaphatidic acid (an activator of Rho) and induced phosphorylation of mitogen-activated protein kinases (Erk1/2, c-jun kinase, and p38). These findings demonstrate that EGCG regulates the structure and growth of HSCs by way of Rho-signaling pathways and suggest that EGCG has therapeutic potential in the setting of liver fibrosis..
99. Rie Sugimoto, Munechika Enjoji, Makoto Nakamuta, Satoshi Ohta, Motoyuki Kohjima, Marie Fukushima, Masami Kuniyoshi, Eiichiro Arimura, Shusuke Morizono, Kazuhiro Kotoh, Hajime Nawata, Effect of IL-4 and IL-13 on collagen production in cultured LI90 human hepatic stellate cells, Liver International, 10.1111/j.1478-3231.2005.01087.x, 25, 2, 420-428, 2005.04, Background: Recently, it has been reported that interleukin 4 (IL-4) and 13 (IL-13) directly activate fibroblasts and promote fibrosis. In the process of hepatic fibrosis, the effects of these cytokines on hepatic stellate cells (HSCs) are not well known. Methods: We evaluated the effects of IL-4 and IL-13 on the collagen production and the proliferation of LI90, a hepatic stellate cell line. We also examined whether interferon (IFN) interferes with the expression of collagen, since IFN has been reported to clinically suppress hepatic fibrosis. Results: The receptor complex for IL-4 and IL-13 was IL-4Rα /IL-13Rα1 on LI90 cells, and the phosphorylation of Stat6 was induced by IL-4 and IL-13. The treatment of LI90 cells with IL-4 or IL-13 increased the production of collagen I protein levels by nearly three times in comparison with untreated cells. Collagen mRNA levels were increased roughly 10-fold by IL-4 and 100-fold by IL-13. Interestingly, BrdU incorporation in LI90 cells was decreased by IL-4 or IL-13 treatment. Furthermore, induction of collagen I production by these cytokines was blocked by IFNα or IFNβ treatment, although neither treatment alone suppressed collagen production. Conclusions: Our data suggested that IL-4 and IL-13 directly affected HSCs by increasing collagen production and suppressing cell proliferation. The anti-fibrogenetic effect of IFN may be due in part to the blockade of IL-4 and IL-13 stimulation of HSCs..
100. Tomoko Izaki, Sachiko Kamakura, Motoyuki Kohjima, Hideki Sumimoto, Phosphorylation-dependent binding of 14-3-3 to Par3β, a human Par3-related cell polarity protein, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2005.01.115, 329, 1, 211-218, 2005.04, Mammalian Par3α and Par3β/Par3L participate in cell polarity establishment and localize to tight junctions of epithelial cells; Par3α acts via binding to atypical PKC (aPKC). Here we show that Par3β as well as Par3α interacts with 14-3-3 proteins in a phosphorylation-dependent manner. In the interaction, Ser-746 of Par3β and the corresponding residue of Par3α (Ser-814) likely play a crucial role, since replacement of these residues by unphosphorylatable alanine results in a loss of interacting activity. The mutant Par3 proteins with the replacement are correctly recruited to tight junctions of MDCK cells and to membrane ruffles induced by an active form of the small GTPase Rac in HeLa cells. Thus, the interaction with 14-3-3 appears to be dispensable to Par3 localization. Consistent with this, the Par3α-14-3-3 interaction does not inhibit the Par3α-aPKC association required for the Par3α localization, although the aPKC-binding site lies close to the Ser-814-containing, 14-3-3-interacting region..
101. Yoshinori Hirano, Sosuke Yoshinaga, Ryu Takeya, Nobuo N. Suzuki, Masataka Horiuchi, Motoyuki Kohjima, Hideki Sumimoto, Fuyuhiko Inagaki, Structure of a cell polarity regulator, a complex between atypical PKC and Par6 PB1 domains, Journal of Biological Chemistry, 10.1074/jbc.M409823200, 280, 10, 9653-9661, 2005.03, A complex of atypical PKC and Par6 is a common regulator for cell polarity-related processes, which is an essential clue to evolutionary conserved cell polarity regulation. Here, we determined the crystal structure of the complex of PKCι and Par6α PB1 domains to a resolution of 1.5 Å. Both PB1 domains adopt a ubiquitin fold. PKCι PB1 presents an OPR, PC. and AID (OPCA) motif, 28 amino acid residues with acidic and hydrophobic residues, which interacts with the conserved lysine residue of Par6α PB1 in a front and back manner. On the interface, several salt bridges are formed including the conserved acidic residues on the OPCA motif of PKCι PB1 and the conserved lysine residue on the Par6α PB1. Structural comparison of the PKCι and Par6α PB1 complex with the p40phox and p67phox PB1 domain complex, subunits of neutrophil NADPH oxidase, reveals that the specific interaction is achieved by tilting the interface so that the insertion or extension in the sequence is engaged in the specificity determinant. The PB1 domain develops the interaction surface on the ubiquitin fold to increase the versatility of molecular interaction..
102. Tsukasa Kawahara, Motoyuki Kohjima, Yuki Kuwano, Hisano Mino, Shigetada Teshima-Kondo, Ryu Takeya, Shohko Tsunawaki, Akihiro Wada, Hideki Sumimoto, Kazuhito Rokutan, Helicobacter pylori lipopolysaccharide activates Rac1 and transcription of NADPH oxidase Nox1 and its organizer NOXO1 in guinea pig gastric mucosal cells, American Journal of Physiology - Cell Physiology, 10.1152/ajpcell.00319.2004, 288, 2 57-2, C450-C457, 2005.02, Primary cultures of guinea pig gastric mucosal cells express NADPH oxidase 1 (Nox1), a homolog of gp91phox, and produce superoxide anion (O 2-) at a rate of ∼100 nmol·mg protein -1· h-1 in response to Helicobacter pylori (H. pylori) lipopolysaccharide (LPS) from virulent type I strains. The upregulated O2- production also enhances H. pylori LPS-stimulated tumor necrosis factor-α or cyclooxygenase-2 mRNA expression, which suggests a potential role for Nox1 in the pathogenesis of H. pylori-associated diseases. The H. pylori LPS-stimulated O2- production in cultured gastric mucosal cells was inhibited by actinomycin D as well as cycloheximide, suggesting that the induction is regulated at the transcriptional level. The LPS treatment not only increased the Nox1 mRNA to a greater extent but also induced expression of the message-encoding, Nox-organizing protein 1 (NOXO1), a novel p47phox homolog required for Nox1 activity. In addition, H. pylori LPS activated Rac1; i.e., it converted Rac1 to the GTP-bound state. A phosphoinositide 3-kinase inhibitor, LY-294002, blocked H. pylori LPS-induced Rac1 activation and O2- generation without interfering with the expression of Nox1 and NOXO1 mRNA. O2 - production inhibited by LY-294002 was completely restored by transfection of an adenoviral vector encoding a constitutively active Rac1 but not an inactive Rac1 or a constitutively active Cdc42. These findings indicate that Rac1 plays a crucial role in Nox1 activation. Thus the H. pylori LPS-stimulated O2- production in gastric mucosal cells appears to require two distinct events: 1) transcriptional upregulation of Nox1 and NOXO1 and 2) activation of Rac1..
103. Munechika Enjoji, E. Arimura, M. Kohjima, K. Kotoh, M. Nakamuta, Urinary N1, N12-diacetylspermine levels correlate with HCV amount in patients with chronic hepatitis C, International Journal of Biological Markers, 10.5301/JBM.2008.1746, 20, 2, 137-138, 2005.01.
104. Kazuhiro Kotoh, Makoto Nakamuta, Motoyuki Kohjima, Marie Fukushima, Shusuke Morizono, Naoya Kobayashi, Munechika Enjoji, Hajime Nawata, Arg-Gly-Asp (RGD) peptide ameliorates carbon tetrachloride-induced liver fibrosis via inhibition of collagen production and acceleration of collagenase activity., International journal of molecular medicine, 14, 6, 1049-1053, 2004.12, Liver cirrhosis is caused by a relative imbalance between synthesis and degradation of collagens. Arg-Gly-Asp (RGD) peptide is a major adhesive domain of several extracellular matrix (ECM) components, such as that involved in the binding of fibronectin to the alpha5beta1 integrin receptor. We previously reported that RGD peptide increased the expression of matrix metalloproteinase in hepatic stellate cells (HSCs) which play a major role in hepatic fibrosis. We evaluated whether RGD-peptides inhibit the progression of liver fibrosis in an animal model of carbon tetrachloride-induced hepatotoxicity. RGD peptide (GRGDS) (1 mg/kg body weight) was injected intraperitoneally (i.p.) 3 times a week for one month. The group treated with control peptide (GRGES) showed pathologically typical hepatic fibrosis, while the RGD-treated group showed minimal fibrotic changes. The liver contents of collagen and hydroxyproline in the RGD-treated group was significantly lower than that of the control group. Collagenase activity measured in liver homogenates was significantly higher in the treated group than in the control group. In an in vitro study using TWNT-4 cells derived from human HSCs, RGD peptide (100 mug/ml) reduced the expression of type I collagen and tissue inhibitor of matrix metalloproteinase-1, and increased that of matrix metalloproteinase-1. These results indicated that RGD peptides inhibited liver fibrosis associated with both decreased collagen production and increased collagenase acitivity, and suggested that RGD peptide might be useful for the therapy of hepatic fibrosis..
105. Shusuke Morizono, Makoto Nakamura, Motoyuki Kohjima, Izuru Miyagi, Tsuyoshi Yoshimoto, Eiichirou Arimaura, Kazuhiro Kotoh, Munechika Enjoji, Yuji Soejima, Akinobu Taketomi, Tomoharu Yoshizumi, Hideaki Uchiyama, Mitsuo Shimada, Yoshihiko Maehara, Hajime Nawata, Evaluation of acute hepatic failure treated at the Department of Medicine III, Kyushu University Hospital
indications for living-donor liver transplantation, Fukuoka igaku zasshi = Hukuoka acta medica, 95, 12, 321-331, 2004.01, To evaluate indications for living-donor liver transplantation (LDLT), we examined 25 consecutive patients with acute hepatic failure admitted to the Department of Medicine III, Kyushu University Hospital between November 2001 and July 2004. These cases were diagnosed as fluminant hepatitis (n=13), severe-type acute hepatitis (n=11), or late-onset hepatic failure (n=1). Nine patients (36%) improved with conservative treatment (conservative treatment group), and the other 16 patients (64%) needed LDLT (LDLT indicated group). In the LDLT indicated group, 11 patients received LDLT, and 4 died because of lack of LDLT donors (n=3), or renal failure (n=1). The LDLT survival rate was 82% (9/11); two patients died due to hepatic infarction and brain edema, respectively. It is very important to predict whether a patient with acute hepatic failure belongs to the conservative treatment group or the LDLT indicated group on admission. Therefore, we analyzed variables that could influence prognosis, including, parameters of hepatic function and platelet counts on admission, and relative hepatic volume (%), which represents the ratio of hepatic volume measured by CT relative to standard hepatic volume calculated with body surface area. Univariate logistic analysis showed that relative hepatic volume, gammaglutamyl transpeptidase (gamma-GTP), alkaline phosphatase (ALP), and the ratio of direct bilirubin to total bilirubin (DB/TB) were significant predictors of survival (p < 0.05). Using these factors plus prothrombin time (PT) and total cholesterol, both of which were relatively significant predictors of survival (p < 0.2), we proposed a model for predicting the probability of survival by the stepwise method. Consequently, we proposed a model using four parameters: ALP, GGTP, PT, and relative hepatic volume (Volume) as shown below: p(%) = 1/(1+exp (-(-36.2375 + ALP x 0.0251 + gamma-GTP x 0.0102 + PT x 0.2558 + Volume 21.2158))) x 100. This model showed a significant correlation between prediction and consequence of survival (r2 = 0.7388, p = 0.0003). In conclusion, LDLT is an effective treatment for acute hepatic failure. The results of this study suggested that our model can adequately predict prognosis in the early phase of acute hepatic failure..
106. Yukiko Noda, Motoyuki Kohjima, Tomoko Izaki, Kazuhisa Ota, Sosuke Yoshinaga, Fuyuhiko Inagaki, Takashi Ito, Hideki Sumimoto, Molecular Recognition in Dimerization between PB1 Domains, Journal of Biological Chemistry, 10.1074/jbc.M306330200, 278, 44, 43516-43524, 2003.10, The PB1 (Phox and Bem 1) domain is a recently identified module that mediates formation of a heterodimeric complex with other PB1 domain, e.g. the complexes between the phagocyte oxidase activators p67phox and p40phox and between the yeast polarity proteins Bem1p and Cdc24p. These PB1 domains harbor either a conserved lysine residue on one side or an acidic OPCA (OPR/PC/AID) motif around the other side; the lysine of p67 phox or Bem1p likely binds to the OPCA of p40phox or Cdc24p, respectively, via electrostatic interactions. To further understand molecular recognition by PB1 domains, here we investigate the interactions mediated by proteins presenting both the lysine and OPCA on a single PB1 domain, namely Par6, atypical protein kinase C (aPKC), and ZIP. Par6 and aPKC form a complex via the interaction of the Par6 lysine with aPKC-OPCA but not via that between the aPKC lysine and Par6-OPCA, thereby localizing to the tight junction of epithelial cells. aPKC also uses its OPCA to interact with ZIP, another protein that has a PB1 domain presenting both the lysine and OPCA, whereas aPKC binds via the conserved lysine to MEK5 in the same manner as ZIP interacts with MEK5. In addition, ZIP can form a homotypic complex via the conserved electrostatic interactions. Thus the PB1 domain appears to be a protein module that fully exploits its two mutually interacting elements in molecular recognition to expand its repertoire of protein-protein interactions..
107. Sosuke Yoshinaga, Motoyuki Kohjima, Kenji Ogura, Masashi Yokochi, Ryu Takeya, Takashi Ito, Hideki Sumimoto, Fuyuhiko Inagaki, The PB1 domain and the PC motif-containing region are structurally similar protein binding modules, EMBO Journal, 10.1093/emboj/cdg475, 22, 19, 4888-4897, 2003.10, The PC motif is evolutionarily conserved together with the PB1 domain, a binding partner of the PC motif-containing protein. For interaction with the PB1 domain, the PC motif-containing region (PCCR) comprising the PC motif and its flanking regions is required. Because the PB1 domain and the PCCR are novel binding modules found in a variety of signaling proteins, their structural and functional characterization is crucial. Bem1p and Cdc24p interact through the PB1-PCCR interaction and regulate cell polarization in budding yeast. Here, we determined a tertiary structure of the PCCR of Cdc24p by NMR. The tertiary structure of the PCCR is similar to that of the PB1 domain of Bem1p, which is classified into a ubiquitin fold. The PC motif portion takes a compact ββα-fold, presented on the ubiquitin scaffold. Mutational studies indicate that the PB1-PCCR interaction is mainly electrostatic. Based on the structural information, we group the PB1 domains and the PCCRs into a novel family, named the PB1 family. Thus, the PB1 family proteins form a specific dimer with each other..
108. Ryu Takeya, Noriko Ueno, Keiichiro Kami, Masahiko Taura, Motoyuki Kohjima, Tomoko Izaki, Hiroyuki Nunoi, Hideki Sumimoto, Novel Human Homologues of p47phox and p67phox Participate in Activation of Superoxide-producing NADPH Oxidases, Journal of Biological Chemistry, 10.1074/jbc.M212856200, 278, 27, 25234-25246, 2003.07, The catalytic core of a superoxide-producing NADPH oxidase (Nox) in phagocytes is gp91phox/Nox2, a membrane-integrated protein that forms a heterodimer with p22phox to constitute flavocytochrome b 558. The cytochrome becomes activated by interacting with the adaptor proteins p47phox and p67phox as well as the small GTPase Rac. Here we describe the cloning of human cDNAs for novel proteins homologous to p47phox and p67phox, designated p41 nox and p51nox, respectively; the former is encoded by NOXO1 (Nox organizer 1), and the latter is encoded by NOXA1 (Nox activator 1). The novel homologue p41nox interacts with p227phox via the two tandem SH3 domains, as does p47phox. The protein p51 nox as well as p67phox can form a complex with p47 phox and with p41nox via the C-terminal SH3 domain and binds to GTP-bound Rac via the N-terminal domain containing four tetratricopeptide repeat motifs. These bindings seem to play important roles, since p47phox and p67phox activate the phagocyte oxidase via the same interactions. Indeed, p41nox and p51nox are capable of replacing the corresponding classical homologue in activation of gp91phox. Nox1, a homologue of gp91phox, also can be activated in cells, when it is coexpressed with p41nox and p51 nox, with p41nox and p67phox, or with p47 phox and p51nox; in the former two cases, Nox1 is partially activated without any stimulants added, suggesting that p41 nox is normally in an active state. Thus, the novel homologues p41nox and p51nox likely function together or in combination with a classical one, thereby activating the two Nox family oxidases..
109. Motoyuki Kohjima, Yukiko Noda, Ryu Takeya, Naoaki Saito, Kosei Takeuchi, Hideki Sumimoto, PAR3β, a novel homologue of the cell polarity protein PAR3, localizes to tight junctions, Biochemical and Biophysical Research Communications, 10.1016/S0006-291X(02)02698-0, 299, 4, 641-646, 2002.12, The cell polarity protein PAR3, conserved from the nematode to the vertebrate, forms a complex with PAR6 and atypical protein kinase C (aPKC), and the protein complex occurs at the tight junctions in mammalian epithelial cells. Here we have cloned human cDNA for a novel PAR3 homologue, designated PAR3β, whose messages are present in a variety of tissues and most abundantly expressed in the adult and fetal kidneys. The encoded protein of 1205 amino acids contains a region homologous to the aPKC-binding domain of PAR3α, another human homologue previously identified, and three PDZ domains; the first PDZ domain of PAR3α is considered to interact with PAR6. Unexpectedly, in contrast to other PAR3s found in various species, PAR3β is incapable of binding to any isotypes of PAR6 or aPKC. Nevertheless PAR3β, expressed intrinsically or extrinsically, localizes to the tight junctions, indicating that the localization does not require the ternary complex formation..