The induction of reactive oxygen species and loss of mitochondrial Omi/HtrA2 is associated with S-nitrosoglutathione-induced apoptosis in human endothelial cells
The induction of reactive oxygen species and loss of mitochondrial Omi/HtrA2 is associated with S-nitrosoglutathione-induced apoptosis in human endothelial cells
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DOI：
10.1016/j.taap.2010.02.004
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发表时间：
2010-05-01
影响因子：
3.8
通讯作者：
Lou, Yi-jia
中科院分区：
文献类型：
--
作者：
Liu, Qi-bing;Liu, Lu-lu;Lou, Yi-jia
S-亚硝基谷胱甘肽（GSNO）诱导的内皮细胞损伤的病理生理相关性尚不清楚。本研究的主要目的是阐明GSNO诱导内皮细胞氧化应激的分子机制。通过DAPI染色和碘化丙啶（PI）流式细胞术检测细胞凋亡的形态学评价。在培养的EA.hy926内皮细胞中，暴露于GSNO导致时间和剂量依赖性凋亡级联反应。当用荧光探针5-（和-6）-羧基-2 '，7'-二氯荧光素二乙酸酯在GSNO处理的细胞中测量细胞内活性氧（ROS）产生时，我们观察到ROS水平升高和伴随的线粒体膜电位损失，表明GSNO诱导的死亡信号传导是通过ROS-线粒体途径介导的。重要的是，我们发现过氧亚硝酸盐的形成和Omi/HtrA 2从线粒体的释放参与了这种现象，而死亡受体依赖性信号的变化在相同的情况下没有检测到。通过药理学方法抑制NADPH氧化酶活化和Omi/HtrA 2提供了针对半胱天冬酶-3活化和GSNO诱导的细胞死亡的显著保护，证实了GSNO以依赖性方式触发内皮细胞中的死亡级联。综上所述，我们的研究结果表明，ROS的过度产生和线粒体Omi/HtrA 2的丢失在活性氮诱导的细胞死亡中起着关键作用，并且这些途径的调节可以具有显著的治疗益处。(C)2010年爱思唯尔公司All rights reserved.
The pathophysiological relevance of S-nitrosoglutathione (GSNO)-induced endothelial cell injury remains unclear. The main objective of this study was to elucidate the molecular mechanisms of GSNO-induced oxidative stress in endothelial cells. Morphological evaluation through DAPI staining and propidium iodide (PI) flow cytometry was used to detect apoptosis. In cultured EA.hy926 endothelial cells, exposure to GSNO led to a time- and dose-dependent apoptotic cascade. When intracellular reactive oxygen species (ROS) production was measured in GSNO-treated cells with the fluorescent probes 5-(and-6)-carboxy-2',7'-dichlorofluorescein diacetate, we observed elevated ROS levels and a concomitant loss in mitochondrial membrane potential, indicating that GSNO-induced death signaling is mediated through a ROS-mitochondrial pathway. Importantly, we found that peroxynitrite formation and Omi/HtrA2 release from mitochondria were involved in this phenomenon, whereas changes of death-receptor dependent signaling were not detected in the same context. The inhibition of NADPH oxidase activation and Omi/HtrA2 by a pharmacological approach provided significant protection against caspase-3 activation and GSNO-induced cell death, confirming that GSNO triggers the death cascade in endothelial cells in a mitochondria-dependent manner. Taken together, our results indicate that ROS overproduction and loss of mitochondrial Omi/HtrA2 play a pivotal role in reactive nitrogen species-induced cell death, and the modulation of these pathways can be of significant therapeutic benefit. (C) 2010 Elsevier Inc. All rights reserved.