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Caveola ATP synthase mediates ATP release in vascular endothelial cells exposed to shear stress

机译:Caveola ATP合成酶在暴露于剪切应力的血管内皮细胞中介导ATP释放

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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. The molecular mechanism of the shear-stress-induced ATP release, however, has not been fully understood. In this study, we have demonstrated that cell-surface ATP synthase is involved in shear stress-induced ATP release. Immuno-fluorescence staining of human pulmonary artery ECs (HPAECs) showed that cell-surface ATP synthase is distributed in lipid rafts and co-localized with caveolin-1. When exposed to shear stress, HPAECs released ATP in a dose-dependent manner, and the ATP release was markedly suppressed by a membrane-impermeable ATP synthase inhibitor, angiostatin, and by an anti-ATP synthase antibody. Depletion of plasma membrane cholesterol with methyl-β cyclo-dextrin (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.
机译:内皮细胞(ECS)释放ATP响应于剪切应力,通过血流量产生的机械力,并且ATP通过激活丙伯对载体释放调节EC功能。然而,剪切应激诱导的ATP释放的分子机制尚未得到完全理解。在该研究中,我们已经证明细胞表面ATP合酶参与剪切应力诱导的ATP释放。人肺动脉ECS(HPAECs)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共同定位。当暴露于剪切应力时,HPAECS以剂量依赖性方式释放ATP,并且通过膜不可渗透的ATP合酶抑制剂,血管抑制素和抗ATP合酶抗体显着抑制ATP释放。具有甲基-β-β-β-糊精(MβCD)的血浆膜胆固醇耗尽破坏了脂质筏并废除了具有Caveolin-1的ATP合酶的共定位,导致剪切应激诱导的ATP释放的显着降低。通过转染Caveolin-1 siRNA的Caveolin-1表达的下调也显着抑制了剪切应力的ATP释放反应。这些结果表明,ATP合酶对Caveolae / Lipid Rafts的定位和靶向对HPAECs的剪切应力诱导的ATP释放至关重要。

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