Protection of Human Umbilical Vein Endothelial Cells against Oxidative Stress by MicroRNA-210.
Protection of Human Umbilical Vein Endothelial Cells against Oxidative Stress by MicroRNA-210.
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MicroRNA-210 保护人脐静脉内皮细胞免受氧化应激。
DOI：
10.1155/2017/3565613
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发表时间：
2017
影响因子：
--
通讯作者：
Liu B
中科院分区：
文献类型：
--
作者：
Li T;Song X;Zhang J;Zhao L;Shi Y;Li Z;Liu J;Liu N;Yan Y;Xiao Y;Tian X;Sun W;Guan Y;Liu B
氧化应激诱导内皮细胞凋亡，促进动脉粥样硬化的发展。MicroRNA-210 （miR-210）与不同细胞类型的凋亡有关。本研究旨在探讨miR-210在氧化应激下人脐静脉内皮细胞（HUVECs）中的作用，并确定其潜在机制。用不同浓度的过氧化氢（H2O2）处理HUVECs，采用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四唑测定法和ATP测定法评估细胞活力。为了评估miR-210在h2o2介导的细胞凋亡中的作用，我们采用了功能获得和功能丧失方法，并使用流式细胞术检测其对细胞凋亡和活性氧（ROS）水平的影响。此外，通过定量逆转录聚合酶链反应（qRT-PCR）检测miR-210的表达，并通过qRT-PCR和Western blot分别在RNA和蛋白水平上评估以下凋亡相关基因的表达：casp8 -相关蛋白2 （CASP8AP2）、caspase-8和caspase-3。结果显示，H2O2以剂量依赖性方式诱导HUVECs细胞凋亡，并增加miR-210表达。在H2O2处理的HUVECs中，过表达miR-210抑制细胞凋亡，降低ROS水平。此外，miR-210在蛋白水平下调CASP8AP2及相关下游caspases。因此，在氧化应激下，miR-210通过减少ROS生成和下调CASP8AP2通路对HUVECs具有促存活和抗凋亡作用。
Oxidative stress induces endothelial cell apoptosis and promotes atherosclerosis development. MicroRNA-210 (miR-210) is linked with apoptosis in different cell types. This study aimed to investigate the role of miR-210 in human umbilical vein endothelial cells (HUVECs) under oxidative stress and to determine the underlying mechanism. HUVECs were treated with different concentrations of hydrogen peroxide (H2O2), and cell viability was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and ATP assay. To evaluate the role of miR-210 in H2O2-mediated apoptosis, gain-and-loss-of-function approaches were used, and the effects on apoptosis and reactive oxygen species (ROS) level were assayed using flow cytometry. Moreover, miR-210 expression was detected by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), and expression of the following apoptosis-related genes was assessed by qRT-PCR and Western blot at the RNA and protein level, respectively: caspase-8-associated protein 2 (CASP8AP2), caspase-8, and caspase-3. The results showed that H2O2 induced apoptosis in HUVECs in a dose-dependent manner and increased miR-210 expression. Overexpression of miR-210 inhibited apoptosis and reduced ROS level in HUVECs treated with H2O2. Furthermore, miR-210 downregulated CASP8AP2 and related downstream caspases at protein level. Thus, under oxidative stress, miR-210 has a prosurvival and antiapoptotic effect on HUVECs by reducing ROS generation and downregulating the CASP8AP2 pathway.