miR-218在高浓度葡萄糖诱导大鼠视网膜微血管内皮细胞凋亡中的作用  

Role of miR-218 in apoptosis of rat retinal microvascular endothelial cells induced by high concentration glucose

作　　者：郭闯 刘学政 GUO Chuang;LIU Xuezheng(The First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121001,Liaoning Province,China;Jinzhou Medical University,Jinzhou 121001,Liaoning Province,China)

机构地区：[1]锦州医科大学附属第一医院急诊外科,辽宁省锦州市121001 [2]锦州医科大学,辽宁省锦州市121001

出　　处：《眼科新进展》2021年第2期116-119,共4页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助项目(编号81571383);辽宁省自然科学基金资助项目(编号2019-ZD-0807)。

摘　　要：目的探讨miR-218在高浓度葡萄糖诱导大鼠视网膜微血管内皮细胞(rat retinal microvascular endothelial cells,rRMECs)凋亡中的作用及其相关机制。方法将rRMECs分为对照组、高糖组、miR-218阴性抑制组和miR-218抑制组。对照组细胞用DMEM低糖培养基培养;高糖组在培养基中添加D-葡萄糖,终浓度为25 mmol·L^(-1);miR-218抑制组及miR-218阴性抑制组是在高糖组培养基的基础上,按照说明书操作,利用Lipo2000将miR-218抑制剂和阴性抑制剂转染rRMECs,共同孵育24 h。提取各组rRMECs总RNA并分析miR-218含量,检测细胞活力,计算细胞凋亡率以及采用Western blot检测各组rRMECs Caspase-3蛋白表达,利用SPSS 19.0统计学软件对实验数据进行单因素方差分析并进行两两比较。结果对照组、高糖组、miR-218阴性抑制组、miR-218抑制组miR-218相对表达量分别为1.00±0.08、1.48±0.12、1.46±0.14、1.03±0.08;细胞活力分别为100.0%、(72.3±7.5)%、(70.7±9.3)%及(98.4±9.4)%;与对照组相比,高糖组和miR-218阴性抑制组的miR-218相对表达量明显升高(均为P<0.05),miR-218抑制组的miR-218相对表达量较对照组变化不显著(P>0.05);高糖组和miR-218阴性抑制组rRMECs细胞活力与对照组相比均明显降低(均为P<0.05),miR-218抑制组与对照组相比细胞活力未见明显变化(P>0.05)。对照组、高糖组、miR-218阴性抑制组、miR-218抑制组rRMECs细胞凋亡率分别为(2.3±0.0)%、(20.3±3.1)%、(23.1±3.2)%及(2.9±0.1)%;高糖组和miR-218阴性抑制组的细胞凋亡率均显著高于对照组(均为P<0.05);而miR-218抑制组与对照组相比,细胞凋亡率变化不明显,差异无统计学意义(P>0.05)。对照组rRMECs Caspase-3蛋白相对表达量为(18.6±2.3)%、高糖组为(43.3±4.5)%、miR-218阴性抑制组为(41.6±3.9)%、miR-218抑制组为(20.0±2.8)%。与对照组相比,高糖组和miR-218阴性抑制组的Caspase-3蛋白相对表达量明显增加(均为P<0.05),miR-218抑制组Caspase-3蛋白相�Objective To explore the role of miR-218 in the apoptosis of rat retinal microvascular endothelial cells(rRMECs)induced by high concentration of glucose and its related mechanisms.Methods The rRMECs were divided into control group,high glucose group,miR-218 negative inhibition group and miR-218 inhibition group.Cells in the control group were cultured with DMEM low-sugar medium;the high glucose group added D-glucose to the medium,and the final concentration was 25 mmol·L^(-1);based on the medium of the high glucose group,Lipo2000 was used to transfect the miR-218 inhibitor and negative control inhibitor into rRMECs following the instructions in the miR-218 inhibition group and miR-218 negative inhibition group for 24 hours.We extracted the total RNA of rRMECs in each group for analyzing the content of miR-218,checking cell viability,and calculating the cell apoptosis rate.Western blot was applied to detect the expression of rRMECs Caspase-3 protein in each group,and SPSS 19.0 statistical software was used to perform one-way analysis of variance on the experimental data for making pairwise comparisons.Results The relative expression levels of miR-218 in the control group,high glucose group,miR-218 negative inhibition group,and miR-218 inhibition group were 1.00±0.08,1.48±0.12,1.46±0.14,1.03±0.08,respectively,and the cell viability was 100.0%,(72.3±7.5)%,(70.7±9.3)%and(98.4±9.4)%,respectively.Compared with the control group,the content of miR-218 in the high glucose group and the miR-218 negative inhibition group increased significantly(both P<0.05).Compared with the control group,the miR-218 content in the miR-218 inhibition group did not change significantly(P>0.05);compared with the control group,the cell viability of rRMECs in the high glucose group and miR-218 negative inhibition group was significantly lower(both P<0.05),compared with the control group,the cell viability of the miR-218 inhibition group did not change significantly(P>0.05).The apoptosis rate of rRMECs in the control group,high glucose

关 键 词：葡萄糖 大鼠视网膜微血管内皮细胞 MIR-218 Caspase-3 细胞凋亡 

分 类 号：R774.1[医药卫生—眼科]