miR-1908抑制高糖诱导的人视网膜血管内皮细胞系HRECs凋亡

目的探讨miR-1908对高糖诱导的人视网膜血管内皮细胞(HRECs)凋亡的影响及分子机制。方法培养HRECs细胞,分为对照组和高糖组,RT-qPCR检测细胞中miR-1908表达水平。转染miR-1908模拟物(mimics)、整合素连接激酶(ILK)的小干扰RNA至HRECs细胞,qRT-PCR和Western blot分别检测miR-1908和ILK蛋白表达验证转染效率。使用高糖干预过表达miR-1908或ILK表达抑制的HRECs细胞,流式细胞仪检测凋亡率,Western blot检测B淋巴细胞瘤-2(Bcl-2)和B淋巴细胞瘤-2相关蛋白(Bax)表达水平。双荧光素酶报告基因实验验证miR-1908和ILK之间关系。结果与对照组比,高糖组HRECs细胞中miR-1908的表达水平显著降低(P<0.05)。过表达miR-1908或ILK表达可降低高糖诱导的HRECs细胞凋亡率(P<0.05),上调Bcl-2蛋白表达(P<0.05),下调Bax蛋白表达(P<0.05)。miR-1908负调控ILK表达,ILK过表达逆转了miR-1908对HRECs细胞凋亡率、Bcl-2和Bax蛋白表达的影响。结论miR-1908可能通过负调控ILK表达上调Bcl-2蛋白表达,下调Bax蛋白表达抑制HRECs细胞的凋亡。

miR-1908 inhibits apoptosis of high glucose-induced human retinal vascular endothelial cell line HRECs

Objective To investigate the effect of miR-1908 on apoptosis of human retinal vascular endothelial cells(HRECs)induced by high glucose concentration and its molecular mechanism.Methods HRECs cells were cultured and divided into normal group and high glucose group.The expression of miR-1908 was detected by RT-qPCR.miR-1908 mimics or small interfering RNA of ILK was transfected into HRECs cells,the expression miR-1908 and ILK protein was detected by RT-qPCR and Western blot in order to detect the transfection efficiency,respectively.Then high concentration of glucose was used to interfere with HRECs with miR-1908 over-expressed or ILK expression inhibited.Apoptosis rate was detected by flow cytometry,Bcl-2 and Bax protein expression was detected by Western blot.The dual luciferase reporter gene assay validated the relationship between miR-1908 and ILK.Results Compared with the control group,the expression level of miR-1908 in HRECs of high glucose group was significantly decreased(P<0.05).miR-1908 over-expressed of or ILK expression decreased the apoptosis rateof HRECs induced by high concentration of glucose(P<0.05),up-regulated Bcl-2 protein expression(P<0.05),and down-regulated Bax protein expression(P<0.05).miR-1908 negatively regulated ILK expression,and over-expression of ILK partially reversed the effect of miR-1908 on apoptosis rate,Bcl-2 and Bax protein expression in HRECs.Conclusions miR-1908 up-regulates the expression of Bcl-2 protein and down-regulates the expression of Bax protein to inhibit the apoptosis of HRECs via negatively regulating ILK expression.