miR-210介导的Bcl-2下调促进动脉粥样硬化内皮细胞凋亡的机制  

Mechanism of Atherosclerosis-Associated Endothelial Cell Apoptosis Promoted by miR-210-Mediated Down-Regulation of Bcl-2

作　　者：任丽 刘婷 吴锡骅 REN Li;LIU Ting;WU Xihua(Department of Neurology,Chancheng District Central Hospital,Foshan 528031,Guangdong,China)

机构地区：[1]禅城区中心医院神经内科,广东佛山528031

出　　处：《贵州医科大学学报》2020年第11期1302-1306,1317,共6页Journal of Guizhou Medical University

基　　金：佛山市科技计划项目(佛科2016AB001521)。

摘　　要：目的:探讨miR-210介导的Bcl-2下调促进动脉粥样硬化内皮细胞凋亡的机制。方法:将常规培养的人主动脉内皮细胞HAECs用100 mg/L氧化低密度脂蛋白(ox-LDL)对细胞进行处理24 h后,分为对照组(Control组)、动脉粥样硬化内皮细胞模型组(ox-LDL组)、miR-210模拟物及抑制剂组;采用CCK-8实验检测细胞活力,细胞凋亡实验检测细胞凋亡水平,qRT-PCR实验检测细胞miR-210的表达,Western blot实验检测细胞Bcl-2蛋白含量,生物信息学算法确定Bcl-2是否为miR-210的潜在靶点,荧光素酶报告基因检测将Bcl-23′-UTR报告质粒(pRL-Bcl-2)转染进修饰过miR-210的HAECs细胞后的荧光素酶活性。结果:经ox-LDL处理过的HAECs细胞的活力低于Control组,凋亡水平高于Control组,miR-210的表达高于Control组,Bcl-2表达低于Control组(P<0.05);miR-210模拟物能抑制Bcl-23′-UTR报告基因的荧光素酶活性(P<0.05),miR-210抑制剂能提高Bcl-23′-UTR报告基因的荧光素酶活性,差异有统计学意义(P<0.05)。结论:miR-210介导的Bcl-2下调促进了动脉粥样硬化内皮细胞的凋亡。Objective:To investigate the mechanism of atherosclerosis-associated endothelial cell apoptosis promoted by miR-210-mediated down-regulation of Bcl-2.Methods:Human aortic endothelial cells(HAECs)treated with 100 mg/L oxidized low density lipoprotein(ox-LDL)for 24 hours were divided into control group,atherosclerosis endothelial cell model group(ox-LDL)and miR-210 simulant and inhibitor group.CCK8 assay was adopted to detect cell viability;apoptosis assay was adopted to detect apoptosis level;qRT-PCR was adopted to detect the expression of miR-210;western blot was adopted to detect the Bcl-2 protein content;bioinformatics algorithm was adopted to determine that Bcl-2 was a potential target of miR-210;and luciferase reporter gene was to detect luciferase activity of Bcl-23'-UTR reporter plasmid(pRL-Bcl-2)transfected into HAECs cells modified with mir-210.Results:The viability level and Bcl-2 expression of HAECs treated with ox-LDL were lower than those of control group;apoptosis level and miR-210 expression of HAECs treated with ox-LDL were higher than those of control group(P<0.05);miR-210 simulant could inhibit the luciferase activity of Bcl-23'-UTR reporter gene(P<0.05);miR-210 inhibitor could increase the luciferase activity of Bcl-23'-UTR reporter gene(P<0.05),among which the differences were statistically significant.Conclusion:miR-210-mediated down-regulation of Bcl-2 promotes the apoptosis of atherosclerotic endothelial cells.

关 键 词：动脉粥样硬化 内皮细胞 细胞凋亡 MIR-210 B细胞淋巴瘤/白血病-2基因Bcl-2 

分 类 号：R54[医药卫生—心血管疾病]