miR-211对高糖诱导下晶状体上皮细胞氧化应激反应的调控作用

目的 探讨miR-211对高糖诱导下晶状体上皮细胞氧化应激反应的调控作用及其可能存在的作用机制.方法 将HLEC-B3细胞在含不同浓度(0、10、20、40 mmol/L)葡萄糖的培养基中培养48 h,RT-PCR检测细胞miR-211表达,Western blot检测细胞SIRT1蛋白表达,试剂盒检测总抗氧化能力(T...

Regulation and mechanism of miR-211 on oxidative stress response of lens epithelial cells induced by high glucose

Objective To investigate the regulatory effect of miR-211 on oxidative stress response of lens epithelial cells induced by high glucose and its possible mechanism. Methods HLEC-B3 cells were cultured in media containing different concentrations of glucose (0 mmol/L, 10 mmol/L, 20 mmol/L, 40 mmol/L) for 48 hours, and the expression of miR-211 was assessed by RT-PCR. SIRT1 protein expression was assessed by Western blot. T-AOC, SOD, GSH-PX and MDA concentrations were assessed by kits. Plasmids containing miR-211 inhibitor and miR-211 NC were transfected into HLEC-B3 cells and cultured in a medium containing 40 mmol/L glucose for 48 h. Concentrations of T-AOC, SOD, GSH-PX, and MDA were assessed by kits, expression of SIRT1 and FoxO3a was assessed by Western blot. Results The expression of miR-211 in HLEB-3 cells was significantly increased with increasing glucose concentration (F=7.271, P<0.001), while the expression of SIRT1 was significantly reduced (F=10.571, P<0.001). The concentration of T-AOC, SOD and GSH-PX in HLEB-3 cells were significantly reduced with the increase of glucose concentration (FT-AOC=10.577, P=0.000; FSOD=12.540, P=0.000; FGSH-PX=15.121, P=0.000), while MDA concentration was significantly increased (FMDA=13.185, P=0.000). The concentrations of T-AOC, SOD, and GSH-PX in miR-211 inhibitor group were significantly higher than those in miR-211 NC group (tT-AOC=5.421, P=0.000; tSOD=10.271, P=0.000; tGSH-PX=7.836, P=0.000), while MDA concentration was significantly lower than that of miR-211 NC group (tMDA=8.757, P=0.000). SIRT1 protein expression in miR-211 inhibitor group was significantly higher than miR-211 NC group (t=15.641, P=0.000), and sFoxO3a protein expression was significantly lower than miR-211 NC group (t=9.823, P=0.000). Conclusion miR-211 can decreased the antioxidative stress capacity-induced by high glucose in lens epithelial cells. This process may be achieved by inhibiting the expression of SIRT1.