Resveratrol regulates mitochondrial reactive oxygen species homeostasis through Sirt3 signaling pathway in human vascular endothelial cells.
Resveratrol regulates mitochondrial reactive oxygen species homeostasis through Sirt3 signaling pathway in human vascular endothelial cells.
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DOI：
10.1038/cddis.2014.530
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发表时间：
2014-12-18
影响因子：
9
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线粒体活性氧（mtROS）稳态在防止内皮细胞氧化损伤中起重要作用，这是动脉粥样硬化形成的初始步骤。白藜芦醇（Resveratrol，RSV）具有多种心脏保护作用，但其对内皮细胞线粒体活性氧稳态的影响尚不清楚。Sirt 3是一种线粒体脱乙酰酶，在线粒体生物能量学中起关键作用，与氧化应激密切相关。本研究的目的是探讨呼吸道合胞病毒（RSV）是否能通过Sirt 3信号通路调节线粒体活性氧（mtROS）稳态来减轻内皮细胞的氧化损伤。我们发现，RSV预处理可以通过增加细胞活力、抑制细胞凋亡、抑制线粒体膜电位崩溃和减少线粒体活性氧的产生来抑制叔丁基过氧化氢（t-BHP）诱导的人脐静脉内皮细胞（HUVEC）氧化损伤。此外，RSV预处理还能提高异柠檬酸脱氢酶2（IDH 2）、谷胱甘肽过氧化物酶（GSH-Px）和锰超氧化物歧化酶（SOD 2）的活性，并使SOD 2脱乙酰化，表明RSV能显著增强t-BHP诱导的内皮细胞对mtROS的清除。同时，RSV通过促进线粒体内Sirt 3富集和随后上调叉头框O3 A（FoxO 3A）介导的ATP 6、CO 1、Cytb、ND 2和ND 5的转录因子编码基因表达，从而导致复合物I活性和ATP合成增加，从而显著减少mtROS的产生。此外，RSV激活磷酸化腺苷单磷酸激活蛋白激酶（p-AMPK）、过氧化物酶体增殖物激活受体γ共激活因子-1 α（PGC-1α）和Sirt 3的表达，以及雌激素相关受体α（ERRα）依赖的Sirt 3 mRNA转录，这些在AMPK抑制剂和AMPK、PGC-1α或Sirt 3 siRNA转染的存在下被消除，说明RSV对线粒体ROS稳态的调节作用依赖于AMPK-PGC-1α-ERRα-Sirt 3信号通路。我们的研究结果表明，一种新的机制，RSV衰减氧化损伤内皮细胞通过调节线粒体ROS稳态，这在一定程度上是通过激活Sirt 3信号通路介导的。
Mitochondrial reactive oxygen species (mtROS) homeostasis plays an essential role in preventing oxidative injury in endothelial cells, an initial step in atherogenesis. Resveratrol (RSV) possesses a variety of cardioprotective activities, however, little is known regarding the effects of RSV on mtROS homeostasis in endothelial cells. Sirt3 is a mitochondrial deacetylase, which plays a key role in mitochondrial bioenergetics and is closely associated with oxidative stress. The goal of the study is to investigate whether RSV could attenuate oxidative injury in endothelial cells via mtROS homeostasis regulation through Sirt3 signaling pathway. We found that pretreatment with RSV suppressed tert-butyl hydroperoxide (t-BHP)-induced oxidative damage in human umbilical vein endothelial cells (HUVECs) by increasing cell viability, inhibiting cell apoptosis, repressing collapse of mitochondrial membrane potential and decreasing mtROS generation. Moreover, the enzymatic activities of isocitrate dehydrogenase 2 (IDH2), glutathione peroxidase (GSH-Px) and manganese superoxide dismutase (SOD2) as well as deacetylation of SOD2 were increased by RSV pretreatment, suggesting RSV notably enhanced mtROS scavenging in t-BHP-induced endothelial cells. Meanwhile, RSV remarkably reduced mtROS generation by promoting Sirt3 enrichment within the mitochondria and subsequent upregulation of forkhead box O3A (FoxO3A)-mediated mitochondria-encoded gene expression of ATP6, CO1, Cytb, ND2 and ND5, thereby leading to increased complex I activity and ATP synthesis. Furthermore, RSV activated the expressions of phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK), peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) and Sirt3, as well as estrogen-related receptor-α (ERRα)-dependent Sirt3 mRNA transcription, which were abolished in the presence of AMPK inhibitor and AMPK, PGC-1α or Sirt3 siRNA transfection, indicating the effects of RSV on mtROS homeostasis regulation were dependent on AMPK-PGC-1α-ERRα-Sirt3 signaling pathway. Our findings indicated a novel mechanism that RSV-attenuated oxidative injury in endothelial cells through the regulation of mtROS homeostasis, which, in part, was mediated through the activation of the Sirt3 signaling pathway.