Shear-stress-induced molecular dynamics of endothelial cell membrane

剪切应力诱导的内皮细胞膜分子动力学

• 批准号: 19300155
• 负责人: YAMAMOTO Kimiko
• 金额: $ 12.15万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2007
• 资助国家: 日本
• 起止时间: 2007 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-19300155/
• 关键词: 血管; 剪断応力; 内皮細胞; カルシウム・シグナリング; メカノトランスダクション; ATP合成酵素; カベオラ; 細胞・組織; 生理学; バイオメカニクス; 機械刺激; ナノイメージング

Endothelial cells (ECs) release ATP in response to shear stress, a mechanical force generated by blood flow, and the ATP released modulates EC functions through activation of purinoceptors. In this study, we have demonstrated that cell-surface ATP synthase is involved in shear-stress-induced ATP release. Immunofluorescence staining of human pulmonary artery ECs (HPAECs) showed that cell-surface ATP synthase is distributed in lipid rafts and co-localized with caveolin-1, a marker protein of caveolae. Immunoprecipitation indicated that the cell-surface ATP synthase and caveolin-1 are physically associated. Measurement of the extracellular metabolism of ^3H-labeled ADP confirmed that cell surface ATP synthase is active in ATP generation. When exposed to shear stress, HPAECs released ATP in a dose-dependent manner, and the ATP release was markedly suppressed by membrane-impermeable ATP synthase inhibitors, angiostatin and piceatannol, and by an anti-ATP synthase antibody. Depletion of plasma membrane cholesterol with methyl-β cyclodextrin (MβCD) disrupted lipid rafts and abolished co-localization of ATP synthase with caveolin-1, which resulted in a marked reduction in shear-stress-induced ATP release. Down-regulation of caveolin-1 expression by transfection of caveolin-1 siRNA also markedly suppressed ATP-releasing responses to shear stress. These results suggest that the localization and targeting of ATP synthase to caveolae/lipid rafts, is critical for shear stress-induced ATP release by HPAECs.

内皮细胞（ECS）响应剪切应力释放ATP，剪切应力，一种由血流产生的机械力，ATP释放的释放通过激活Purinoceptor的EC功能调节EC功能。在这项研究中，我们证明了细胞表面ATP合酶参与剪切压力诱导的ATP释放。人肺动脉ECS（HPAEC）的免疫荧光染色表明，细胞表面ATP合酶分布在脂质筏中，并与Caveolin-1共定位，Caveolin-1是小窝的标记蛋白。免疫沉淀表明细胞表面ATP合酶和小窝蛋白1与物理相关。测量 ^3H标记ADP的细胞外代谢证实了细胞表面ATP合酶在ATP生成中活跃。当暴露于剪切应力时，HPAECS以剂量依赖性的方式释放ATP，并且ATP释放被膜上可侵蚀的ATP合酶抑制剂，Angiostatin和piceatannol以及抗-ATP合酶抗体明显抑制。用甲基-β环糊精（MβCD）消耗质膜胆固醇（MβCD）破坏了脂质筏，并消除了Caveolin-1的ATP合酶的共定位，从而导致剪切压力诱导的ATP释放显着减少。通过转染小窝蛋白-1 siRNA对小窝蛋白-1表达的下调也显着抑制了对剪切应力的释放反应。这些结果表明，将ATP合酶定位和靶向口腔/脂质筏，对于HPAECS剪切应力诱导的ATP释放至关重要。

项目成果

流れ剪断応力はVEGF-Notchシグナルを介して血管内皮前駆細胞を動脈化する

流动剪切应力通过 VEGF-Notch 信号使血管内皮祖细胞动脉化

• 发表时间: 2009
• 作者: 桝村智美、山本希美子;他3名
• 通讯作者: 他3名

流れずり応力が惹起する内皮細胞膜特性の変化

流动剪切应力引起的内皮细胞膜特性的变化

• 发表时间: 2009
• 作者: 山本希美子;神谷瞭;安藤譲二
• 通讯作者: 安藤譲二

血流による血管トーヌス調節因子P2×4

血流诱发血管紧张度调节因子P2×4

• 发表时间: 2007
• 期刊: 医学のあゆみ(総説) 223
• 作者: 安藤 譲二;山本 希美子
• 通讯作者: 山本 希美子

Up-regulation of cell surface P2X4 receptor by flow shear stress

流动剪切应力上调细胞表面 P2X4 受体

• 发表时间: 2008
• 作者: 小林剛;山本希美子 他
• 通讯作者: 山本希美子 他

Endothelial P2X4-mediated shear-stress-mechanotransduction and its roles in the control of vascular functions Fukuoka Purine 2009

内皮 P2X4 介导的剪切应力机械转导及其在血管功能控制中的作用福冈嘌呤 2009

• 发表时间: 2009
• 作者: K. Yamamoto;J. Ando
• 通讯作者: J. Ando