高糖环境下mircoRNA-200b影响视网膜血管内皮细胞功能的机制研究

Effect of micro RNA-200b on function of retinal endothelial cells under high-glucose environment and related mechanism

目的探讨高糖环境下micro RNA-200b(mi R-200b)影响人视网膜血管内皮细胞(h RECs)功能的作用机制。方法在正常和高糖环境下培养h RECs,实时荧光定量聚合酶链式反应检测mi R-200b表达。分别用mi R-200b和特异性mi R-200b抑制剂转染h RECs,并且用噻唑蓝法检测高糖环境下mi R-200b对h RECs增殖的影响。用Western blot测定h RECs转染mi R-200b和mi R-200b抑制剂后对血管内皮生长因子(VEGF)、转化生长因子-β1(TGF-β1)蛋白和m RNA的表达的影响。结果高糖组VEGF、TGF-β1 m RNA和蛋白表达较对照组增加(P<0.05)。转染mi R-200b后,h RECs中mi R-200b表达升高,而VEGF、TGF-β1 m RNA和蛋白的表达降低。与高糖组比较,h RECs增殖受到抑制(P<0.05)。结论 mi R-200b通过改变VEGF和TGF-β1的表达,调节h RECs生长和增殖,延缓糖尿病视网膜病变的发生、发展。

Objective To investigate the effect of mleroRNA-200b （miR-200b） on human retinal endothe- lial cells （hRECs） under high-glucose environment and its mechanism. Methods hRECs were cultured under high-glucose or normal environment. Real time PCR was applied to detect miR-200b expression, miR-200b was transfected to hRECs and MTT was used to detect its effect on hRECs proliferation under high-glucose environment. Real time PCR and Western blot were performed to determine VEGF and TGF-β1 expressions in the retina endothelial cells. Results Compared with the normal control group, VEGF and TGF-β1 mRNA and protein expressions increased markedly in the high-glucose group （P〈 0.05）. After miR-200b transfection, miR-200b expression increased in the hRECs, while VEGF and TGF-β1 mRNA and protein expressions decreased obviously. Compared with the high-glucose group, hRECs proliferation was inhibited （P 〈 0.05）. Conclusions miR-200b can regulate the growth and proliferation of RECs by changing VEGF and TGF-β1 expression and delay diabetic retinopathy.