Bim介导氧化应激诱导的人晶状体上皮细胞凋亡

Apoptosis mediated by Bim of human lens epithelial cells induced by oxidative stress

目的:研究Bim在氧化应激诱导的人晶状体上皮细胞凋亡中的作用。方法:过氧化氢(H2O2)刺激体外培养的人晶状体上皮细胞,Hoechst33258染色细胞核,计数细胞凋亡率;western blot检测Bim及Caspase-3蛋白表达水平;RT-PCR检测BimmRNA表达水平;小分子干扰片段敲除Bim后观察干扰效率。结果:H2O2刺激人晶状体上皮细胞4、8、12h后,凋亡率显著增加,与对照组相比有显著差异(P<0.05)。Bim mRNA及蛋白质呈时间依赖的表达上调。小分子干扰Bim的表达后,细胞12h凋亡率从约(49.91±1.20)%下降到约(14.23±0.90)%(siBim1)、18.44%±1.52%(siBim2)。结论:Bim介导了H2O2诱导的人晶状体上皮细胞凋亡,可能在白内障的发生发展中发挥重要作用。

Objective To investigate the role of Bim play in the apoptosis of human lens epithelial cells （HLEC） induced by oxidative stress. Methods HLEC which were cultured in vitro were treated with H202, and then the apoptosis was quantified by counting the cells with pyknotic nuclei after staining with Hoechst33258. The expressions of Bim and Caspase-3 protein were detected by western blot, and the expression of Bim mRNA was detected by RT-PCR. The apoptosis was observed after Bim was knocked down. Results After being treated with n202 for 4 h, 8 h, and 12 h, the apoptosis rates of HLEC were significantly increased than those in control group （P 〈 0.05）. Up-regulation of Bim mRNA and protein was induced by H202 in a time-dependent manner. The knockdown of Bim expression could decrease the apoptosis rates from （49.91±1.20）% to （14.23±0.90）% （siBiml） and （ 18.44±1.52）% （siBim2） at 12 h. Conclusion The apoptosis of HLEC induced by H202 was mediated by Bim, which may play an important role in the development of cataract.