九州大学 研究者情報
研究者情報 (研究者の方へ)入力に際してお困りですか?
基本情報 研究活動 教育活動 社会活動
平野 真弓(ヒラノ マユミ) データ更新日:2019.06.17

助教 /  医学研究院 基礎医学部門 生体情報科学


主な研究テーマ
血管内皮細胞の細胞間接触による増殖停止機構の解明。
キーワード:血管内皮細胞、細胞間接触、増殖停止、細胞周期、転写調節、
1997.04~2021.03.
プロテイナーゼ活性化型受容体1を介する血管機能障害の分子機構解明
キーワード:血管内皮細胞、プロテイナーゼ活性化受容体1、血管機能障害、血管透過性亢進
2015.04~2021.03.
トロンビンによる内皮細胞活性化機構の解明
キーワード:血管内皮細胞、ミオシン軽鎖のリン酸化、ストレスファイバー、バリアー機能
2010.04~2021.03.
トロンビン受容体の活性制御異常の分子機構解明と新たな動脈硬化治療戦略の開発
キーワード:血管内皮細胞、トロンビン、プロテイナーゼ活性化型受容体、血管透過性、動脈硬化、NO
2012.04.
血管内皮細胞の細胞間接触による増殖停止における細胞周期制御因子p27Kip1の転写調節機構の解明。
キーワード:血管内皮細胞、細胞間接触、増殖停止、細胞周期、転写調節、Rac1
2001.04~2010.03.
血管内皮細胞のトロンビンによる新たなNO産生機構の解明。
キーワード:血管内皮細胞、プロテイナーゼ活性化型受容体、トロンビン、NO、Ca
2003.04~2012.03.
内皮細胞の貯蔵部作動性Ca流入機構に関する研究
キーワード:血管内皮細胞、カルシウム流入、カルシウム貯蔵部、
2007.04.
従事しているプロジェクト研究
プロテイナーゼ活性化型受容体1を介する血管機能障害の分子機構解明
2015.04~2015.06, 代表者:平野真弓, 九州大学大学院医学研究院 分子細胞情報学, 日本.
トロンビン受容体の活性制御異常の分子機構解明と新たな動脈硬化治療戦略の開発
2012.04~2015.03, 代表者:平野真弓, 九州大学大学院医学研究院
虚血性心臓病、脳梗塞などの血管病の克服には、動脈硬化の予防と治療が最重要課題となる。血栓-血管壁相互作用を担うトロンビン受容体は、動脈硬化の病態形成に重要な役割を果たす。申請者は、これまでの研究から「トロンビン受容体の活性制御異常(=発現亢進と脱感作障害)」が血管攣縮などの血管病の病態形成に重要な役割を果たすことを見出した。
また、トロンビンが引き起こす血管内皮の透過性の亢進にミオシン軽鎖の2リン酸化が重要な役割を果たすことを見出した。
本研究は、動脈硬化の新たな治療標的と期待されるトロンビン受容体の観点から、従来の予防、治療法にはない新たな戦略の開発を目指す。.
トロンビン受容体を標的とする核心的脳血管攣縮治療法の開発
2014.04~2016.03, 代表者:平野勝也, 香川大学.
肺高血圧症におけるプロテイナーゼ活性化型受容体の役割解明と治療応用
2011.04~2014.03, 代表者:平野勝也, 九州大学大学院医学研究院
肺高血圧症は、肺血管抵抗が進行性に上昇し、最終的に右心不全を来して死に至る重篤な疾患である。種々の薬物治療が行われているにも関わらず、5年生存率は未だ40%に留まる。新たな作用機序に基づく治療法の開発が急務である。本研究では、トロンビンに対する肺動脈の反応特性に関する独自の知見を基に新たな肺高血圧治療法の開発に挑む。これまでの研究から、肺高血圧症は、脳血管攣縮と並び、トロンビン受容体標的薬の治療効果が期待される。肺高血圧患者の病理像を再現する独自開発の動物モデルを用いて、ヒトの病態に即して研究を行い、トロンビン受容体の新たな視点から従来にない治療法を開発する点が本研究の特徴である。.
血管病変形成初期における内皮細胞プロテイナーゼ活性化型受容体の役割
2008.04~2011.03, 代表者:平野真弓, 九州大学大学院医学研究院
血管機能に重大な影響を及ぼす、凝固系と血管壁細胞の相互作用を仲介する最も重要な仕組であるプロテイナーゼ活性化型受容体 (proteinase-activated receptor; PAR) は比較的最近同定された受容体であり、内皮細胞の活性化や透過性亢進におけるPARの役割やメカニズムには未だ不明な点が多い。本研究では、PARという新しい視点から、凝固系と内皮細胞の相互作用の仕組みを分子レベルで明らかにし、動脈硬化病変形成の初期過程を理解し、新たな治療標的を確立することを目的とする。.
細胞間接触による内皮細胞の増殖と機能分化に関わるシグナル伝達網の解明
2005.04~2008.03, 代表者:平野真弓, 九州大学大学院医学研究院
血管内皮細胞は、細胞間接触により増殖を停止すると機能分化する。一方、内皮細胞が傷害を受け細胞間接触が破綻すると、再び増殖性を取り戻し、機能異常を来すため、動脈硬化等の血管病の成因となる。これまでに細胞間接触による増殖停止の際、細胞周期制御因子p27Kip1タンパク質の発現が亢進すること、この発現亢進は転写の活性化とmRNAの安定化によることを明らかにした。本研究では、p27Kip1の転写調節領域や転写因子を明らかにし、細胞間接触による増殖停止・分化の仕組みを分子レベルで明らかにすることを目的とする。.
細胞間接触による血管内皮細胞の増殖停止に関わるシグナル伝達網の解明
2002.04~2005.03, 代表者:平野真弓, 九州大学大学院医学研究院
血管内皮細胞は、細胞間接触により増殖を停止すると機能分化し、血管の緊張と増殖の維持に重要な役割を果たす。一方、接触による増殖抑制が破綻すると内皮細胞の脱分化を引き起こす。本研究では、「細胞間接触の受容」から「核における細胞周期制御」に至る分子機構を明らかにすることをの目的とした。.
宇宙環境利用に関する地上研究
2001.04~2004.03, 代表者:平野勝也, 財団法人日本宇宙フォーラム(日本)
血管の構築と機能制御に及ぼす重力変動の影響をあきらかにする。.
研究業績
主要著書
1. Hirano K, Hirano M, Abe S and Kanaide H., Cytosolic calcium transients in vascular smooth muscle. in Ion Channels of Vascular Smooth Muscle Cells and Endothelial Cells, eds Sperelakis N and Kuriyama H., Elsivier, New York,, 93-105,, 1991.01.
主要原著論文
1. Mayumi Hirano, Katsuya Hirano, Myosin di-phosphorylation and peripheral actin bundle formation as initial events during endothelial barrier disruption , Scientific Report , 10.1038/srep20989, 6: 20989, 2016.02, The phosphorylation of the 20-kD myosin light chain (MLC) and actin filament formation play a key role in endothelial barrier disruption. MLC is either mono- or di-phosphorylated (pMLC and ppMLC) at T18 or S19. The present study investigated whether there are any distinct roles of pMLC and ppMLC in barrier disruption induced by thrombin. Thrombin induced a modest bi-phasic increase in pMLC and a robust mono-phasic increase in ppMLC. pMLC localized in the perinuclear cytoplasm during the initial phase, while ppMLC localized in the cell periphery, where actin bundles were formed. Later, the actin bundles were rearranged into stress fibers, where pMLC co-localized. Rho-kinase inhibitors inhibited thrombin-induced barrier disruption and peripheral localization of ppMLC and actin bundles. The double, but not single, mutation of phosphorylation sites abolished the formation of peripheral actin bundles and the barrier disruption, indicating that mono-phosphorylation of MLC at either T18 or S19 is functionally sufficient for barrier disruption. Namely, the peripheral localization, but not the degree of phosphorylation, is suggested to be essential for the functional effect of ppMLC. These results suggest that MLC phosphorylation and actin bundle formation in cell periphery are initial events during barrier disruption. .
2. Toshiro Saito, mayumi hirano, Kenji Sunagawa, Katsuya Hirano, Pivotal Role of Rho-Associated Kinase 2 in Generating the Intrinsic Circadian Rhythm of Vascular Contractility. , Circulation 127(1), 10.1161/CIRCULATIONAHA.112.135608 , 127, 104-114, 2013.01, Background—The circadian variation in the incidence of cardiovascular events may be attributable to the circadian changes in vascular contractility. The circadian rhythm of vascular contractility is determined by the interplay between the central and peripheral clocks. However, the molecular mechanism of the vascular intrinsic clock that generates the circadian rhythm of vascular contractility still remains largely unknown.
Methods and Results—The agonist-induced phosphorylation of myosin light chain in cultured smooth muscle cells synchronized by dexamethasone pulse treatment exhibited an apparent circadian oscillation, with a 25.4-hour cycle length. The pharmacological inhibition and knockdown of Rho-associated kinase 2 (ROCK2) abolished the circadian rhythm of myosin light chain phosphorylation. The expression and activity of ROCK2 exhibited a circadian rhythm in phase with that of myosin light chain phosphorylation. A clock gene, RORα, activated the promoter of the ROCK2 gene, whereas its knockdown abolished the rhythmic expression of ROCK2. In the mouse aorta, ROCK2 expression exhibited the circadian oscillation, with a peak at Zeitgeber time 0/24 and a nadir at Zeitgeber time 12. The myofilament Ca2+ sensitization induced by GTPγS and U46619, a thromboxane A2 analog, at Zeitgeber time 0/24 was greater than that seen at Zeitgeber time 12. The circadian rhythm of ROCK2 expression and myofilament Ca2+ sensitivity was abolished in staggerer mutant mice, which lack a functional RORα.
Conclusions—ROCK2 plays a pivotal role in generating the intrinsic circadian rhythm of vascular contractility by receiving a cue from RORα. The ROCK2-mediated intrinsic rhythm of vascular contractility may underlie the diurnal variation of the incidence of cardiovascular diseases..
3. Yuichiro Kikkawa, S Matsuo, Katsuharu Kameda, Katsuya Hirano, A Nakamizo, mayumi hirano, Tomio Sasaki, Mechanisms underlying potentiation of endothelin-1-induced myofilament Ca2+ sensitization after subarachnoid hemorrhage.
, J Cereb Blood Flow Metab , 32, 341-352, 2012.03, Increased vascular smooth muscle contractility has an important role in the development of cerebral vasospasm after subarachnoid hemorrhage (SAH). Myofilament Ca2+ sensitivity is a major determinant of smooth muscle contractility. We investigated changes in the Ca2+-sensitizing effect of endothelin-1 (ET-1) and the mechanisms underlying ET-1-induced Ca2+ sensitization after SAH using a rabbit SAH model. After SAH, the contractile response to ET-1 was enhanced, and the ETA receptor expression was upregulated in the basilar artery. In a-toxin-permeabilized preparations, ET-1 induced enhanced and prolonged contraction after SAH, suggesting that ET-1-induced Ca2+ sensitization is potentiated after SAH. Endothelin-1-induced Ca2+ sensitization became less sensitive to inhibitors of Rho-associated coiled-coil protein kinase (ROCK) and protein kinase C (PKC) after SAH. The expression of PKCa, ROCK2, PKC-potentiated phosphatase inhibitor of 17 kDa (CPI-17) and myosin phosphatase target subunit 1 (MYPT1) was upregulated, and the level of phosphorylation of CPI-17 and MYPT1 was elevated after SAH. This study demonstrated for the first time that the Ca2+-sensitizing effect of ET-1 on myofilaments is potentiated after SAH. The increased expression and activity of PKCa, ROCK2, CPI-17, and MYPT1, as well as the upregulation of ETA receptor expression are suggested to underlie the enhanced and prolonged Ca2+sensitization induced by ET-1..
4. Katsuharu Kameda, Yuichiro Kikkawa, mayumi hirano, S Matsuo, Tomio Sasaki, Hideo Kanaide, Combined argatroban and anti-oxidative agents prevents increased vascular contractility to thrombin and other ligands after subarachnoid hemorrhage2+-sensitivity and induces vasoconstriction in porcine pulmonary arteries., Br J Pharmacol, 165 , 106-119, 2012.01.
5. Aman M, Hirano M, Kanaide H, Hirano K, Up-regulation of proteinase-activated receptor-2 and increased response to trypsin in endothelial cells after exposure to oxidative stress in rat aortas., J Vasc Res, 47, 494-506, 2010.10.
6. Hirano K, Hirano M, Hanada A:, Involvement of STIM1 in the proteinase-activated receptor 1-mediated Ca2+ influx in vascular endothelial cells., J Cell Biochem, 108, 499-507, 2009.09.
7. Maeda Y, Hirano K, Hirano M, Kikkawa Y, Kameda K, Sasaki T, Kanaide H, Enhanced contractile response of the basilar artery to platelet-derived growth factor in subarachnoid hemorrhage., Stroke 40, 591-596, 2009, 40, 591-596, 2009.02.
8. Hirano M, Kanaide H, Hirano K, Rac1-dependent transcriptional up-regulation of p27Kip1 by homophilic cell-cell contact in vascular endothelial cells., Biochim Biophys Acta -Mol Cell Res 1773, 1500-1510, 2007, 1773, 1500-1510, 2007
, 2007.08.
9. Yufu T, Hirano K, Bi D, Hirano M, Nishimura J, Iwamoto Y, Kanaide H, Rac1 regulation of the surface expression of PAR1 and responsiveness to thrombin in vascular smooth muscle cells, Arterioscler Thromb Vasc Biol, 10.1161/01.ATV.0000168418.10276.f0, 25, 7, 1506-1511, vol. 25, 1506-1511, 2005.01.
10. Shiga N, Hirano K, Hirano M, Nishimura J, Nawata H, Kanaide H, Long-term inhibition of RhoA attenuates vascular contractility by enhancing endothelial NO production in an intact rabbit mesenteric artery, Cir Res, 10.1161/01.RES.0000165483.34603.91, 96, 9, 1014-1021, vol. 96, 1014-1021, 2005.01.
11. Koga M, Hirano K, Hirano M, Nishimura J, Kanaide H:, Akt plays a central role in the anti-apoptotic effect of estrogen in endothelial cells, Biochem Biophys Res Commun, 10.1016/j.bbrc.2004.09.060, 324, 1, 321-325, vol. 324, 321-325, 2004.01.
12. Hirano K, Ihara E, Hirano M, Nishimura J, Nawata H, Kanaide H:, Facilitation of proteasomal degradation of p27Kip1 by N-terminal cleavage, and their sequence requirements, FEBS Lett, 10.1016/j.febslet.2004.08.014, 574, 1-3, 111-115, vol. 574, 111-115, 2004.01.
13. Hirano K, Hirano M, Nishimura J, Kanaide H:, A critical period requiring Rho proteins for cell cycle progression uncovered by reversible protein transduction in endothelial cells, FEBS Lett, 10.1016/j.febslet.2004.05.084, 570, 1-3, 149-154, vol. 570, 149-154, 2004.01.
14. Nakayama T, Hirano K, Hirano M. Nishimura J, Kuga H, Nakamura K, Takahashi S, Kanaide H:, Inactivation of protease-activated receptor-1 by proteolytic removal of the ligand region in vascular endothelial cells, Biochem Pharmacol, 10.1016/j.bcp.2004.03.005, 68, 1, 23-32, vol. 68, 23-32, 2004.01.
15. Hirano K, Derkach DN, Hirano M, Nishimura J, Takahashi S, Kanaide H, Transduction of the N-terminal fragments of MYPT1 enhances myofilament Ca2+ sensitivity in an intact coronary artery, Arterioscler Thromb Vasc Biol, 10.1161/01.ATV.0000116028.42230.4c, 24, 3, 464-469, vol. 24, 464-469, 2004.01.
16. Eto W, Hirano K, Hirano M, Nishimura J, Kanaide H:, Intracellular alkalinization induces Ca2+ influx via non-voltage-operated Ca2+ channels in the rat aortic smooth muscle cells, Cell Calcium, 10.1016/S0143-4160(03)00151-9, 34, 6, 477-484, vol. 34, 477-484, 2003.01.
17. Hirano K, Zeng Y, Hirano M, Nishismura J, Kanaide H, Sequence requirement for nuclear localization and growth inhibition of p27Kip1R, a degradation-resistant isoform of p27Kip1, J Cell Biochem, 10.1002/jcb.10499, 89, 1, 191-202, vol. 89, 191-202, 2003.01.
18. Ihara E, Hirano K, Hirano M, Nishimura J, Nawata H, Kanaide H, The mechanism of down-regulation of L-type Ca2+ channel in the proliferating smooth muscle cells of rat aorta, J Cell Biochem 87, 242-251, vol. 87, 242-251, 2002.01.
19. Hirano M, Hirano K, Nishimura J, Kanaide H, Transcriptional up-regulation of p27Kip1 during contact-induced growth arrest in vascular endothelial cells, Exp Cell Res, 10.1006/excr.2001.5384, 271, 2, 356-367, vol. 271, 356-367, 2001.01.
20. Hirano K, Hirano M, Zeng Y, Nishimura J, Hara K, Muta K, Nawata H, Kanaide H, Cloning and functional expression of a degradation-resistant novel isoform of p27Kip1, Biochem J, 10.1042/0264-6021:3530051, 353, 51-57, vol. 353, 51-57, 2001.01.
21. Hirano K, Hirano M, Eto W, Nishimura J, Kanaide H, Mitogen-induced upregulation of non-smooth muscle isoform of a-tropomyosin in rat aortic smooth muscle cells, Eur J Pharmacol, 10.1016/S0014-2999(00)00681-6, 406, 2, 209-218, vol. 406, 209-218, 2000.01.
22. Hirano M, Niiro N, Hirano K, Nishimura J, Hartshorne DJ, Kanaide H: , Expression, subcellular localization and cloning of the 130 kDa regulatory subunit of myosin phosphatase in porcine aortic endothelial cells.
, Biochem. Biophys. Res. Commun. , 10.1006/bbrc.1998.9973, 254, 2, 490-496, 1999.01.
主要総説, 論評, 解説, 書評, 報告書等
1. Mayumi Hirarno, 平野 勝也, 平成20年度〜22年度科学研究費補助金・基盤研究(C) 研究成果報告書
「血管病変形成初期における内皮細胞プロテイナーゼ活性化型受容体の役割」
, 2011.05.
2. Mayumi Hirarno, 平野 勝也, 平成16年度〜17年度科学研究費補助金・基盤研究(C)(2) 研究成果報告書
「細胞間接触による内皮細胞の増殖と機能分化に関わるシグナル伝達網の解明」
, 2006.03.
3. Mayumi Hirarno, 平野 勝也, シグマ アルドリッチ ジャパン株式会社 SIGMA 抗体カタログVol.2、pp 193, 701
「ブタ大動脈内皮細胞のβカテニン免疫蛍光染色写真」提供
, 2004.05.
4. Mayumi Hirarno, 平野 勝也, 平成14年度〜15年度科学研究費補助金・基盤研究(C)(2) 研究成果報告書、
「細胞間接触による血管内皮細胞の増殖停止に関わる細胞内シグナル伝達網の解明」
, 2004.03.
主要学会発表等
1. Mayumi Hirano, Katsuya Hirano, Submembranous di-phosphorylation of myosin light chain and actin fiber formation play a critical role as an initial event during endothelial barrier disruption, 11th International Symposium on Resistance Arteries ISRA, 2014.09, The phosphorylation of myosin light chain (MLC) plays an essential role in endothelial barrier disruption. MLC is phosphorylated at Thr18 and Ser19. We elucidated distinct role of mono- and di-phosphorylation of MLC during barrier disruption in porcine aortic endothelial cells. Thrombin decreased trans-endothelial electrical resistance (TEER) with a bottom at 3-5 min. Phos-tag SDS-PAGE analysis revealed that 25% and 2% of MLC were mono- and di-phosphorylated (pMLC and ppMLC) before stimulation. pMLC slightly increased in the cytoplasmic region 3 min after stimulation, while ppMLC increased to 35% in the submembranous region, where actin fiber formation was also induced. Thereafter, ppMLC returned to the resting level, while pMLC reached a second heave after 15 min, when stress fiber formation was observed. Rho-kinase inhibitor abolished thrombin-induced decrease in TEER, with inhibition of the submembranous ppMLC and actin fiber formation at 3 min as well as pMLC at 15 min. Myosin ATPase inhibitor, blebbistatin, inhibited the decrease in TEER with no effect on MLC phosphorylation. Adenoviral expression of a mutant of MLC with substitution of Thr18 and Ser19 to Ala (MLCT18A+S19A) abolished the thrombin-induced submembranous ppMLC and actin fiber formation as well as the decrease in TEER. However, those events were observed in the cells expressing MLCT18A or MLCS19A as in the cells expressing wild-type MLC. In conclusion, the submembranous ppMLC and actin fiber formation and the resultant circumferential contraction serve as an initial event during endothelial barrier disruption. Mono-phosphorylation of MLC is functionally sufficient for submembranous actin fiber formation and barrier disruption..
2. 平野 真弓, 平野 勝也, Role of mono- and di-phosphorylation of myosin light chain in thrombin-induced endothelial barrier disruption., 第87回日本薬理学会, 2014.03.
3. 平野 真弓, 平野 勝也, 細胞膜直下におけるミオシン軽鎖2リン酸化反応とアクチン線維束形成が、血管内皮バリアー障害の初期事象として重要な役割を果たす, 第4回福岡薬理・生理学研究会, 2013.12.
4. 平野 真弓, 平野 勝也, 花田亜希子, 血管内皮細胞における小胞体カルシウムセンサー蛋白質STIM1 の役割, 生理研研究会2013 , 2013.11.
5. Mayumi Hirano, Katsuya Hirano, Di-phosphorylation of myosin light chain and actin fiber formation in the submembranous region at inter-endothelial junction play a crucial role in thrombin-induced endothelial barrier disruption, American Heart Association Scientific Session, 2013, 2013.11.
6. Mayumi Hirano, Katsuya Hirano, Myosin light chain di-phosphorylation at sub-membranous region plays a critical role during thrombin-induced barrier disruption in vascular endothelial cells., International Symposium on Regulatory circuits in cell motility, honoring Dave Hartshorne, 2013.10.
7. 平野 真弓, 平野 勝也, 血管平滑筋細胞トロンビン受容体の脱感作障害のメカニズム, 第55回日本平滑筋学会総会, 2013.08.
8. Katsuya Hirano, mayumi hirano, Critical Role of Rho-kinase-mediated Subjunctional Di-phosphorylation of Myosin Light Chain (MLC) in Thrombin-induced Disruption of Endothelial Barrier, 第 77 回日本循環器学会学術集会, 2013.03.
9. Katsuya Hirano, Katsuharu Kameda, Akiko Hanada, mayumi hirano, Oxidative stress and Extracellular Signal-regulated Kinase (ERK) Play a Key Role in Impairment of Receptor Desensitization and Prolongation of Vascular Reactivity in the Cerebral Artery after Subarachnoid Hemorrhage., International Stroke Conference 2013, 2013.02.
10. mayumi hirano, Akiko Hanada, Katsuya Hirano, Rho Kinase-mediated Di-phosphorylation of Myosin Light Chain in the Sub-membranous Regions and Circumferential Actomyosin Contraction Mediate Thrombin-induced Barrier Disruption in Vascular Endothelial Cells., International Stroke Conference 2013, 2013.02.
11. Mayumi hirano, Katsuya Hirano, 血管平滑筋細胞におけるトロンビン受容体の脱感作障害にExtracellular signal-regulated kinase (ERK) が重要な役割を果たす, 第65回日本薬理学会西南部会, 2012.11.
12. mayumi hirano, Akiko Hanada, Katsuya Hirano, Oxidative Stress and Rho-associated Coiled-coil Protein Kinase (ROCK)-mediated Double Phosphorylation of Myosin Light Chain in the Submembranous Region Plays a Key Role in Thrombin-induced Barrier Dysfunction in Vascular Endothelial Cells, Scientific Sessions 2012 of the American Heart Association, 2012.11.
13. Toshiro Saito, mayumi hirano, Tomomi Ide, Toshihiro Ichiki, Noriyuki Koibuchi, Kenji Sunagawa, Katsuya Hirano, A Clock Gene ROR-mediated Regulation of the Activity of Rho-associated Coiled-coil Protein Kinase 2 (ROCK2) Plays a Key Role in Generating Vascular Intrinsic Circadian Rhythm of Myofilament Ca2+ Sensitivity and Vascular Contractility, Scientific Sessions 2012 of the American Heart Association, 2012.11.
14. mayumi hirano, Katsuya Hirano, Di-phosphorylation of myosin light chain (MLC) in the sub-membranous region plays an important role in the thrombin-induced barrier dysfunction in endothelial cells, 第29回国際心臓研究学会(ISHR)日本部会総会, 2012.10.
15. mayumi hirano, Akiko Hanada, Katsuya Hirano, Rho kinase-mediated di-phosphorylation of myosin light chain in the subjunctional regions plays a critical role in thrombin-induced endothelial barrier dysfunction, 第85回 日本薬理学会, 2012.03.
16. Katsuharu kameda, Yuichiro Kikkawa, mayumi hirano, O Matsuo, Tomio Sasaki, Katsuya Hirano, Oxidative Stress Impairs Desensitization of the G-protein Coupled Receptors (GPCRs) and Increases Vascular Reactivity by Prolonging the Contractile Responses in the Cerebral Arteries after Subarachnoid Hemorrhage., Scientific Sessions 2011 of the American Heart Association, 2011.11.
17. mayumi hirano, Akiko Hanada, Katsuya Hirano, Depletion of Intracellular Ca2+ Stores Induces Phosphorylation of Stromal Interaction Molecular 1 (STIM1), which Contributes to the Sustained Phase of Store-operated Ca2+ Influx in Vascular Endothelial Cells
, Scientific Sessions 2011 of the American Heart Association, 2011.11.
18. Hirano K,Akiko Hanada, Hirano M, Ca2+-independent Di-phosphorylation of Myosin Light Chain by Rho-associated Coiled-coil Protein Kinase (ROCK) in the Subjunctional Regions of Cell-Cell Contact Mediates Thrombin-induced Barrier Dysfunction in Vascular Endothelial Cells, Scientific Sessions 2011 of the American Heart Association, 2011.11, A disruption of endothelial barrier function is a predisposing factor for the development of atherosclerosis. The phosphorylation of myosin light chain (MLC) is a key signal of barrier dysfunction. MLC phosphorylation is thought to take place sequentially at S19 and then T18, induces actin stress fiber formation and generates traction force to disrupt cell-cell contact. However, it is unclear how the phosphorylation at two sites contributes to barrier dysfunction. The present study investigated the role of mono- and di-phosphorylation of MLC (MLC-P and MLC-PP) in thrombin-induced barrier dysfunction. Thrombin (1 u/mL) increased the cytosolic Ca2+ concentration and decreased the transendothelial electrical resistance (TEER) with a peak at 1 and 3-5 min, respectively, in porcine aortic endothelial cells (PAEC). A new SDS-PAGE method using a Phos-tagTM compound, which specifically binds to a phosphate group, allowed separate quantification of MLC-P and MLC-PP. PAEC at confluence contained 25% MLC-P and 2% MLC-PP under resting conditions. Thrombin stimulation only slightly increased MLC-P at 3 min, while it showed up on stress fibers. MLC-PP increased to a peak of 35% at 3-5 min, and thereafter declined to the resting level within 15 min. MLC-PP was localized in the subjunctional region of cell-cell contact in addition to the stress fibers at 3 min. ROCK inhibitors, Y27632 and H1152, inhibited the thrombin-induced decrease in TEER, the increase in MLC-PP and the subjunctional localization of MLC-PP, while having no effect on the Ca2+ response. The MLC-P level remained unaffected by Y27632, while the stress fiber localization was inhibited. Gq inhibitor, YM254890, inhibited the Ca2+ response and the decrease in TEER, but it had no effect on the increase in MLC-PP. The introduction of dominant negative mutants of G12, G13 and Gq using a cell-penetrating peptide inhibited the thrombin-induced decrease in TEER. The present study provides the first evidence that MLC-P and MLC-PP are differentially regulated in endothelial cells, and that Ca2+-independent, ROCK-mediated MLC-PP in the subjunctional regions plays a critical role in the thrombin-induced barrier dysfunction. Inhibition of MLC-PP is thus necessary to restore endothelial barrier dysfunction..
19. Hirano K, Hanada A, Hirano M, Store-operated phosphorylation of endogenous STIM1 in the interphase cells as revealed by a Phos-tag SDS-PAGE analysis., 第84回日本薬理学会, 2011.03.
20. Kameda K, Kikkawa Y, Hirano M, Sasaki T, Hirano K, The combination of argatroban and vitamin C normalizes the increased vascular reactivity after subarachnoid hemorrhage(SAH)., 第84回日本薬理学会, 2011.03.
21. Hirano M,Hanada A, Hirano K, Thrombin causes barrier dysfunction via two distinct signal transduction pat hways in vascular
endothelial cells., 第84回日本薬理学会, 2011.03.
22. 亀田勝治、吉川雄一郎、平野真弓、平野勝也, アルガトロバンとビタミンCの併用はくも膜下出血後のトロンビンに対する収縮反応性亢進を予防する, 第63回日本薬理学会西南部会、第20回日韓薬理学合同セミナー, 2010.11.
23. 平野勝也、平野真弓、花田亜希子, Stromal interaction molecule 1 (STIM1)のリン酸化による貯蔵部作動性カルシウム流入の制御, 第63回日本薬理学会西南部会、第20回日韓薬理学合同セミナー, 2010.11.
24. Hirano M, Hanada A, Hirano K, Involvement of two signaling pathways in the thrombin- induced barrier dysfunction in vascular endothelial cells., 第63回日本薬理学会西南部会、第20回日韓薬理学合同セミナー, 2010.11.
25. Saito T, Hirano M, Ide T, Sunagawa K, Hirano K, Identification of Rho-associated kinase 2 (ROCK2) as a key molecule generating the intrinsic circadian rhythm of vascular contractility. , Scientific Sessions 2010 of the American Heart Association, 2010.11.
26. Hirano K, Maki J, Hirano M, Rho-associated kinase (ROCK) mediates the thrombin-induced production of reactive oxygen species and myosin light chain phosphorylation-independent vasoconstriction in the pulmonary artery.. , Scientific Sessions 2010 of the American Heart Association, 2010.11.
27. Saito T, Hirano M, Ide T, Sunagawa K, Hirano K , Rho kinase plays a key role in the vascular intrinsic clock system that generates the circadian change in smooth muscle contractility. 9th International Conference of the Cardiovascular System Dynamics Society, September 23-26, 2010, Fukuoka, Japan, 9th International Conference of the Cardiovascular System Dynamics Society, 2010.09.
28. 平野勝也、平野真弓, 血管内皮細胞のストア作動性Ca2+流入の調節と生理的役割, 第87回日本生理学会大会シンポジウム「血管トーヌス調節の分子機構とその異常, 2010.05.
29. 吉川雄一郎、亀田勝治、平野真弓、佐々木富男、平野勝也, くも膜下出血後の脳血管収縮性亢進メカニズム, 第87回日本生理学会大会シンポジウム「血管トーヌス調節の分子機構とその異常, 2010.05.
30. Saito T, Hirano M, Ide T, Sunagawa K, Hirano K , Intrinsic circadian oscillation of myosin light chain phosphorylation in vascular smooth muscle cells. , Experimental Biology 2010, 2010.04.
31. Saito T, Hirano M, Hirano K, ROCK2 plays a key role in the circadian change in thrombin-induced myosin light chain Phosphorylation in smooth muscle. (横浜、2010.3. 22), 第84回日本薬理学会, 2010.03.
32. Kameda K, Kikkawa Y, Hirano M, Sasaki T, Hirano K, The combination of thrombin inhibitor and vitamin C normalizes the increased vascular reactivity after subarachnoid hemorrhage., 第83回日本薬理学会, 2010.03.
33. Saito T, Hirano M, Hirano K, The thrombin-induced myosin light chain phosphorylation exhibits an intrinsic circadian oscillation in vascular smooth muscle., 第83回日本薬理学会, 2010.03.
34. Hirano M, Hirano K, Differential regulation of the thrombin-induced mono- and di-phosphorylation of myosin light chain in vascular endothelial cells., 第83回日本薬理学会, 2010.03.
35. Kikkawa Y, Kameda K, Hirano M, Sasaki T, Hirano K, Impaired feedback regulation of the receptor signaling and the myofilament Ca2+ sensitization contributes to increased vascular reactivity after subarachnoid hemorrhage., 第83回日本薬理学会, 2010.03.
36. Hirano K, Hanada A, Hirano M, Intracellular alkalinization activates STIM1-dependent but store-independent Ca2+ influx in vascular endothelial cells., 第83回日本薬理学会, 2010.03.
37. Hirano K, Hirano M, Stromal interaction molecule-1 (STIM1) plays an important role in the thrombin-induced Ca2+ signal and Ca2+-dependent NO production in endothelial cells., 第74回日本循環器学会総会, 2010.03.
38. Kikkawa Y, Kameda K, Hirano M, Hirano K, Impaired feedback inhibition of the receptor signaling and the myofilament Ca2+ sensitization contributes to increased vascular reactiveness after subarachnoid hemorrhage., 第74回日本循環器学会総会, 2010.03.
39. Kameda K, Kikkawa Y, Hirano M, Hirano K, Combination of thrombin inhibitor and antioxidative agent provides a novel strategy to normalize the increased vascular reactivity after subarachnoid hemorrhage., 第74回日本循環器学会総会, 2010.03.
40. 平野勝也、平野真弓、花田亜希子, トロンビンが引き起こす血管内皮細胞のCa2+シグナルとNO産生におけるStromal interaction molecule 1 (STIM1)の関与, 第62回日本薬理学会西南部会, 2009.11.
41. 吉川雄一郎、亀田勝治、平野真弓、平野勝也, くも膜下出血ウサギ脳底動脈における受容体活性および収縮装置のカルシウム感受性に対するフィードバック調節機構の障害。, 第62回日本薬理学会西南部会, 2009.11.
42. Maki J, Hirano M, Hirano K:, Thrombin activation of proteinase-activated receptor 1 (PAR1) induces Ca2+-independent vasoconstriction by activating the production of reactive oxygen species in the pulmonary artery. , Scientific Sessions 2009 of the American Heart Association, 2009.11.
43. Mayumi Hirano, Katsuya Hirano, Phos-tagTM SDS-PAGE analysis revealed differential requirement of Ca2+ source for the thrombin-induced mono- and di-phosphorylation of myosin light chain in vascular endothelial cells., 国際、シンポジウム Satellite Symposium of the IUPS 2009 , Post-genomic Advances in the Physiology of Smooth Muscle, 2009.07.
44. Hirano K, Hirano M, Reactive oxygen species mediates thrombin-induced noncanonical vasoconstriction independent of Ca2+ and myosin light chain phosphorylation in pulmonary artery, 第73回日本循環器学会総会, 2009.03.
45. Hirano K, Hirano M, Enhanced contractile response of the basilar artery to platelet-derived growth factor in subarachnoid hemorrhage, 第73回日本循環器学会総会, 2009.03.
46. Hirano M、Hirano K, A new method for analyzing myosin light chain phosphorylation with a Phos-tagTM technology., 第82回日本薬理学会, 2009.03.
47. Kikkawa Y, Kameda K, Hirano M, Hirano K, The impairment of the desensitization of G protein-coupled receptors in subarachnoid hemorrhage in rabbit basilar arteries., 第82回日本薬理学会, 2009.03.
48. Kameda K, Kikkawa Y, Hirano M, Hirano K, Preventive effect of thrombin inhibitor argatroban on increased contractile response to thrombin in the basilar artery after subarachnoid hemorrhage., 第82回日本薬理学会, 2009.03.
49. Shiga K, Hirano M, Hirano K, Roles of proteinase-activated receptors in the increased detrusor muscle contractility and pollakisuria in cystitis in mice., 第82回日本薬理学会, 2009.03.
50. Hirano K, Hanada A, Hirano M, Involvement of STIM1 in PAR1-mediated Ca2+ influx and NO production in vascular endothelial cells., 第82回日本薬理学会, 2009.03.
51. Maki J, Hirano M, Hirano K, Roles of reactive oxygen species (ROS) and Rho-kinase in the thrombin-induced and proteinase-activated receptor 1 (PAR1)-mediated pulmonary artery vasoconstriction., 第82回日本薬理学会, 2009.03.
52. Hirano K, Hanada A, Hirano M, Identification of a novel STIM1-binding protein as a regulator of store-operated Ca2+ influx., 第82回日本薬理学会, 2009.03.
53. 平野勝也、牧盾、平野真弓, トロンビンが引き起こす、カルシウム及びミオシンリン酸化非依存性肺動脈収縮反応における活性酸素の役割。, 第61回日本薬理学会西南部会, 2008.11.
54. 平野勝也、牧盾、平野真弓, トロンビンが引き起こす肺動脈収縮反応におけるRhoキナーゼの多面的役割, 第61回日本薬理学会西南部会, 2008.11.
55. 平野真弓、平野勝也, トロンビンが引き起こす血管内皮細胞のCa2+流入におけるSTIM1の関与, トランスポーターワークショップIN福岡, 2008.11.
特許出願・取得
特許出願件数  1件
特許登録件数  1件
学会活動
所属学会名
米国心臓協会
日本循環器学会
日本生理学会
日本薬理学会
日本分子生物学会
日本平滑筋学会
日本心脈管作動物質学会
学協会役員等への就任
2013.02~2015.02, 第44回日本心脈管作動物質学会, 組織委員.
2004.03, 日本薬理学会, 評議員.
学会大会・会議・シンポジウム等における役割
2014.03.19~2014.03.21, 第88回日本薬理学会年会, 座長(Chairmanship).
2015.02.06~2015.02.07, 日本心脈管作動物質学会, 組織委員.
学術論文等の審査
年度 外国語雑誌査読論文数 日本語雑誌査読論文数 国際会議録査読論文数 国内会議録査読論文数 合計
2016年度      
2014年度      
2008年度      
その他の研究活動
海外渡航状況, 海外での教育研究歴
Universitu of Arizona, UnitedStatesofAmerica, 1990.10~1996.11.
研究資金
科学研究費補助金の採択状況(文部科学省、日本学術振興会)
2015年度~2017年度, 基盤研究(C), 代表, プロテイナーゼ活性化型受容体1を介する血管機能障害の分子機構解明.
2014年度~2015年度, 萌芽研究, 分担, トロンビン受容体を標的とする核心的脳血管攣縮治療法の開発.
2012年度~2014年度, 基盤研究(C), 代表, トロンビン受容体の活性制御異常の分子機構解明と新たな動脈硬化治療戦略の開発.
2011年度~2013年度, 基盤研究(C), 分担, 肺高血圧症におけるプロテイナーゼ活性化型受容体の役割解明と治療応用
.
2008年度~2010年度, 基盤研究(C), 代表, 血管病変形成初期における内皮細胞プロテイナーゼ活性化型受容体の役割.
2002年度~2003年度, 基盤研究(C), 代表, 細胞間接触による血管内皮細胞の増殖停止に関わる細胞内シグナル伝達網の解明.
2004年度~2005年度, 基盤研究(C), 代表, 細胞間接触による内皮細胞の増殖と機能分化に関わるシグナル伝達網の解明.
2005年度~2006年度, 基盤研究(C), 分担, 血管病変部の緊張異常と増殖性亢進におけるプロテイナーゼ活性化型受容体の役割.
競争的資金(受託研究を含む)の採択状況
2009年度~2009年度, ライフサイエンス振興財団 平成22年度 研究開発の助成, 分担, くも膜下出血後の脳血管攣縮と神経障害に対するトロンビン受容体を標的とした新たな治療戦略の開発.
2009年度~2009年度, 平成21年度財団法人宮田心臓病研究振興基金, 分担, 「肺高血圧におけるトロンビン受容体の病態生理学的役割の解明と新たな治療法の開発」.
2008年度~2008年度, 財団法人 武田科学振興財団・2008年度「報彰基金」研究奨励金, 分担, 「くも膜下出血後脳血管攣縮の分子機構解明と新たな予防・治療法の開発」.
2007年度~2009年度, 横山藤逸臨床薬理研究助成基金・平成19年度第1回医療研究助成, 分担, プロテイナーゼ活性化型受容体及びその発現調節機構を標的とした血管病治療の新戦略.
2007年度~2007年度, , 分担, Development of Novel Therapeutic Strategies Targeting Proteinase-Activated Receptors for the Prevention and Treatment of Atherothrombotic Vascular Diseases.

九大関連コンテンツ

pure2017年10月2日から、「九州大学研究者情報」を補完するデータベースとして、Elsevier社の「Pure」による研究業績の公開を開始しました。
 
 
九州大学知的財産本部「九州大学Seeds集」