Li Tianyi, Song Xianjing, Zhang Jichang, Zhao Lei, Shi Yongfeng, Li Zhibo, Liu Jia, Liu Ning, Yan Youyou, Xiao Yanlong, Tian Xin, Sun Wei, Guan Yinuo, Liu Bin
Department of Cardiology, The Second Hospital of Jilin University, Changchun, Jilin, China.
Oxid Med Cell Longev. 2017;2017:3565613. doi: 10.1155/2017/3565613. Epub 2017 Mar 7.
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 (HO), 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 HO-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 HO 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 HO. 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.
氧化应激诱导内皮细胞凋亡并促进动脉粥样硬化发展。微小RNA-210（miR-210）与不同细胞类型的凋亡相关。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（HO）处理HUVECs，采用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐法和ATP法评估细胞活力。为了评估miR-210在HO介导的凋亡中的作用，采用了功能获得和功能缺失方法，并使用流式细胞术检测对凋亡和活性氧（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表达，并分别通过qRT-PCR和蛋白质印迹在RNA和蛋白质水平评估以下凋亡相关基因的表达：半胱天冬酶-8相关蛋白2（CASP8AP2）、半胱天冬酶-8和半胱天冬酶-3。结果表明，HO以剂量依赖性方式诱导HUVECs凋亡并增加miR-210表达。miR-210过表达抑制了HO处理的HUVECs的凋亡并降低了ROS水平。此外，miR-210在蛋白质水平下调了CASP8AP2和相关下游半胱天冬酶。因此，在氧化应激下，miR-210通过减少ROS生成和下调CASP8AP2途径对HUVECs具有促生存和抗凋亡作用。
