Molecular mechanisms of angiotensin II-mediated mitochondrial dysfunction - Linking mitochondrial oxidative damage and vascular endothelial dysfunction
Molecular mechanisms of angiotensin II-mediated mitochondrial dysfunction - Linking mitochondrial oxidative damage and vascular endothelial dysfunction
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DOI：
10.1161/circresaha.107.162800
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发表时间：
2008-02-29
影响因子：
20.1
通讯作者：
Dikalov, Sergey I.
中科院分区：
文献类型：
--
作者：
Doughan, Abdulrahman K.;Harrison, David G.;Dikalov, Sergey I.
线粒体功能障碍是大多数心血管疾病的显着特征。血管紧张素 (Ang) II 是动脉粥样硬化和高血压的重要刺激因素；然而，它对线粒体功能的影响仍然未知。我们假设 Ang II 可以诱导线粒体氧化损伤，进而可能降低内皮一氧化氮（NO 中心点）的生物利用度并促进血管氧化应激。使用电子自旋共振、二氢乙锭高效液相色谱法、Amplex Red 和阳离子染料荧光，研究了 Ang II 对分离线粒体和完整牛主动脉内皮细胞中线粒体 ROS、线粒体呼吸、膜电位、谷胱甘肽和内皮 NO 中心点的影响。 Ang II 显着增加线粒体 H2O2 的产生。将完整细胞与罗布麻素（NADPH 氧化酶抑制剂）、尿酸（过氧亚硝酸盐清除剂）、白屈菜红碱（蛋白激酶 C 抑制剂）、N-G-硝基-L-精氨酸甲酯（一氧化氮合酶抑制剂）、5-羟基癸酸酯（线粒体 ATP 敏感剂）预孵育可阻断这种增加。 钾通道抑制剂）或格列本脲。用小干扰 RNA 消耗 NADPH 氧化酶的 p22(phox) 亚基也抑制了 Ang II 介导的线粒体 ROS 产生。 Ang II 耗尽线粒体谷胱甘肽，增加状态 4 呼吸并减少状态 3 呼吸，并降低线粒体呼吸控制比。这些反应被罗布麻宁、5-羟基癸酸酯和格列本脲减弱。此外，5-羟基癸酸可防止 Ang II 诱导的内皮 NO 中心点和线粒体膜电位下降。因此，Ang II 通过激活内皮细胞 NADPH 氧化酶和过氧亚硝酸盐的形成，通过蛋白激酶 C 依赖性途径诱导线粒体功能障碍。此外，响应 Ang II 的线粒体功能障碍调节内皮 NO 中心点和条形上方的 O-2（中心点）（-）。的产生，进而对内皮功能障碍的发展产生影响。
Mitochondrial dysfunction is a prominent feature of most cardiovascular diseases. Angiotensin (Ang) II is an important stimulus for atherogenesis and hypertension; however, its effects on mitochondrial function remain unknown. We hypothesized that Ang II could induce mitochondrial oxidative damage that in turn might decrease endothelial nitric oxide (NO center dot) bioavailability and promote vascular oxidative stress. The effect of Ang II on mitochondrial ROS, mitochondrial respiration, membrane potential, glutathione, and endothelial NO center dot was studied in isolated mitochondria and intact bovine aortic endothelial cells using electron spin resonance, dihydroethidium high-performance liquid chromatography-based assay, Amplex Red and cationic dye fluorescence. Ang II significantly increased mitochondrial H2O2 production. This increase was blocked by preincubation of intact cells with apocynin (NADPH oxidase inhibitor), uric acid (scavenger of peroxynitrite), chelerythrine (protein kinase C inhibitor), N-G-nitro-L-arginine methyl ester (nitric oxide synthase inhibitor), 5-hydroxydecanoate (mitochondrial ATP-sensitive potassium channels inhibitor), or glibenclamide. Depletion of p22(phox) subunit of NADPH oxidase with small interfering RNA also inhibited Ang II-mediated mitochondrial ROS production. Ang II depleted mitochondrial glutathione, increased state 4 and decreased state 3 respirations, and diminished mitochondrial respiratory control ratio. These responses were attenuated by apocynin, 5-hydroxydecanoate, and glibenclamide. In addition, 5- hydroxydecanoate prevented the Ang II-induced decrease in endothelial NO center dot and mitochondrial membrane potential. Therefore, Ang II induces mitochondrial dysfunction via a protein kinase C-dependent pathway by activating the endothelial cell NADPH oxidase and formation of peroxynitrite. Furthermore, mitochondrial dysfunction in response to Ang II modulates endothelial NO center dot and O-2(center dot)(-) over bar. generation, which in turn has ramifications for development of endothelial dysfunction.