miR-193a-3p抑制高糖诱导人视网膜血管内皮细胞凋亡

miR-193a-3p inhibits high glucose-induced apoptosis of human retinal vascular endothelial cells

目的探讨miR-193a-3p对高糖诱导的人视网膜血管内皮细胞(HRECs)凋亡的影响和分子机制。方法体外培养HRECs细胞,建立高糖(葡萄糖浓度为30 mmol/L)HRECs细胞模型。设置对照组、模型组、anti-miR-NC、ani-miR-193a-3p组、模型+miR-NC组、模型+miR-193a-3p组、模型+LY294002。RT-qPCR检测miR-193a-3p的表达水平;Western blot检测活化的含半胱氨酸的天冬氨酸蛋白水解酶3(c-caspase-3)、B细胞淋巴瘤/白血病-2(Bcl-2)、磷酸化的磷酸肌醇3激酶(p-PI3K)和磷酸化的蛋白激酶B(p-AKT)的表达水平;流式细胞仪检测细胞凋亡。结果模型组与对照组比较,miR-193a-3p、Bcl-2表达显著降低,凋亡率、c-caspase-3、p-PI3K和p-AKT表达显著升高(P<0.05)。低表达miR-193a-3p促进c-caspase-3、p-PI3K和p-AKT表达,抑制Bcl-2表达,促进细胞凋亡(P<0.05)。高表达miR-193a-3p可减轻高糖处理对HRECs凋亡、c-caspase-3和Bcl-2表达的影响(P<0.05)。抑制PI3K/AKT信号通路可减轻高糖处理对HRECs凋亡、c-caspase-3和Bcl-2表达的影响(P<0.05)。结论miR-193a-3p可能通过调控PI3K/AKT信号通路抑制高糖诱导HRECs凋亡。

Objective To investigate the effect of miR-193a-3p on apoptosis induced by high glucose in human retinal vascular endothelial cells(HRECs)and its molecular mechanism.Methods HRECs cells were cultured in vitro to establish a high glucose(30 mmol/L glucose concentration)cell model.HRECs were divided into control group,model group,anti-miR-NC group,anti-miR-193a-3p group,model+miR-NC group,model+miR-193a-3p group,and model+LY294002 group.RT-qPCR was used to detect the expression of miR-193a-3p;Western blot was used to detect the expression levels of cleaved-caspase-3(c-caspase-3),B cell lymphoma/leukemia-2(Bcl-2),phosphorylated phosphoinositide 3 kinase(p-PI3K)and phosphorylated protein kinase B(p-AKT);flow cytometry was used to detect cell apoptosis.Results Compared with the control group,the expressions of miR-193a-3p and Bcl-2 in the model group was significantly decreased,whereas apoptosis rate,and the expres-sion of c-caspase-3,P-PI3K and P-Akt was significantly increased(P<0.05).Low expression of miR-193a-3p promotes c-caspase-3,p-PI3K and p-AKT expression,inhibited Bcl-2 expression,and promoted cell apoptosis(P<0.05).High expression of miR-193a-3p can significantly reduce the effect of high glucose treatment on the apoptosis,c-caspase-3 and Bcl-2 expression of HRECs(P<0.05).Inhibition of PI3K/AKT signaling pathway reversed the effect of high glucose treatment the apoptosis,c-caspase-3 and Bcl-2 expression of HRECs(P<0.05).Conclusions miR-193a-3p could inhibit high glucose-induced apoptosis of HRECs by regulating PI3K/AKT signaling pathway.