MiR-3202对高糖诱导的人视网膜血管内皮细胞损伤的影响  

The effect of miR-3202 on human retinal vascular endothelial cell injury induced by high glucose

作　　者：罗红 袁昌亮[1] 陈岚[1] Luo Hong;Yuan Changliang;Chen Lan(Department of Ophthalmology,Hubei Maternal and Child Health Hospital,430070 Wuhan,China)

机构地区：[1]湖北省妇幼保健院眼科,武汉430070

出　　处：《中华细胞与干细胞杂志（电子版）》2021年第3期155-160,共6页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：湖北省卫健委基金(JX4C34)。

摘　　要：目的探讨微小RNA-3202(miR-3202)对高糖诱导人视网膜血管内皮细胞(HRCEC)凋亡的影响及其潜在的分子机制。方法体外培养HRCEC,分为低糖组、高糖组、高糖+miR-NC组、高糖+miR-3202组、高糖+si-NC组、高糖+si-ILK组、高糖+miR-3202+pcDNA3.1组和高糖+miR-3202+pcDNA3.1-ILK组。采用实时荧光定量PCR(qRT-PCR)与Western blot法测定HRCEC中miR-3202、整合素连接激酶(ILK)的表达;流式细胞术测定细胞凋亡率;双荧光素酶报告基因实验与Western blot实验验证miR-3202与ILK的靶向调控作用;Western blot测定HRCEC中Bax和Bcl-2蛋白的表达。采用两样本t检验进行统计学分析。结果与低糖组比较,高糖组HRCEC中miR-3202的表达水平(0.95±0.09比0.21±0.02)降低,ILK的表达水平(0.30±0.03比0.86±0.08)、细胞凋亡率(8.54﹪±1.03﹪比28.53﹪±3.25﹪)均升高,差异具有统计学意义(P均<0.05);与高糖+miR-NC组比较,高糖+miR-3202组细胞凋亡率(28.86﹪±2.45﹪比12.67﹪±1.15﹪)、Bax的表达水平(1.12±0.11比0.36±0.03)均下降,Bcl-2的表达水平(0.23±0.02比0.77±0.07)升高,差异具有统计学意义(P均<0.05);与高糖+si-NC组比较,高糖+si-ILK组细胞凋亡率(28.86﹪±2.45﹪比13.46﹪±1.24﹪)、Bax的表达水平(1.10±0.11比0.47±0.05)均降低,Bcl-2的表达水平(0.23±0.02比0.64±0.06)上升,差异具有统计学意义(P均<0.05);miR-3202可负向调控靶基因ILK的表达,ILK过表达可逆转miR-3202对高糖诱导的HRCEC凋亡的作用。结论miR-3202可通过下调ILK的表达而抑制高糖诱导的HRCEC凋亡。Objective To investigate the effect of microRNA-3202(miR-3202)on apoptosis induced by high glucose in human retinal vascular endothelial cells(HRCEC)and its molecular mechanisms.Methods HRCEC was cultured in vitro and divided into eight groups,including low glucose group,high glucose group,high glucose+miR-NC group,high glucose+miR-3202 group,high glucose+si-NC group,high glucose+si-ILK group,high sugar+miR-3202+pcDNA3.1 group,high glucose+miR-3202+pcDNA3.1-ILK group.The expression of miR-3202 and ILK in HRCEC was determined by real-time quantitative PCR(qRT-PCR)and Western blot,and the apoptotic rate was determined by flow cytometry.The dual luciferase reporter gene assay and Western blot assay were used to verified the relationship between miR-3202 and ILK.Also,The expression of Bax and Bcl-2 protein in HRCEC was determined by Western blot.Results Compared with the low glucose group,the expression level of miR-3202 in the HRCEC of the high glucose group was significantly decreased(0.95±0.09 vs 0.21±0.02,P<0.05),and the expression level of ILK(0.30±0.03 vs 0.86±0.08)and the rate of cell apoptosis were significantly increased(8.54﹪±1.03﹪vs 28.53﹪±3.25﹪,P<0.05).Compared with the high glucose+miR-NC group,the apoptotic rate of the high glucose+miR-3202 group(28.86﹪±2.45﹪vs 12.67﹪±1.15﹪)and the expression level of Bax were significantly decreased(1.12±0.11 vs 0.36±0.03,P<0.05),however,the expression level of Bcl-2 significantly increased(0.23±0.02 vs 0.77±0.07,P<0.05).Compared with the high glucose+si-NC group,the apoptotic rate of the high glucose+si-ILK group(28.86﹪±2.45﹪vs 13.46﹪±1.24﹪,P<0.05)and the expression level of Bax were both significantly decreased(1.10±0.11 vs 0.47±0.05,P<0.05),while the expression level of Bcl-2 was significantly increased(0.23±0.02 vs 0.64±0.06,P<0.05).MiR-3202 may negatively regulate the expression of the target gene ILK.Overexpression of ILK reversed the effect of miR-3202 on high glucose-induced apoptosis of HRCEC.Conclusion high glucose ca

关 键 词：miR-3202 整合素连接激酶 糖尿病视网膜病变 凋亡 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]