miR-34a/SIRT1在H2O2诱导人晶状体上皮细胞氧化应激中的表达

目的 观察微小RNA-34a(microRNA-34a,miR-34a)和沉默信息调节因子1(silent information regulator 1,SIRT1)在不同浓度H2O2诱导的人晶状体上皮细胞(SRA01/04细胞)氧化应激中的表达变化.方法 用不同浓度的H2O2(0μmol·L-1、100 μmol·L-1、200 μmol·L-1、300 μmol·L-1、400 μmol·L-1)处理细胞24 h后,用CCK-8检测细胞存活率,流式细胞仪检测细胞凋亡率和RT-PCR检测miR-34a/SIRT1的表达.结果 CCK-8检测结果显示:100～400 μmol·L-1 H2O2对SRA01/04细胞有增殖抑制作用,且呈剂量依赖关系,与0μmol·L-1 H202组相比差异均有统计学意义(均为P＜0.01);流式细胞仪凋亡检测结果显示:0μmol·L-1 H2O2组细胞凋亡率为(6.1±1.2)％;100 μmol·L-1、200μmol·L-1、300 μmol·L-1 H2O2组细胞凋亡率分别为(26.3±1.8)％、(32.5±2.2)％、(64.7±5.3)％,各组与0μmol·L-1H2O2组比较,差异均有统计学意义(均为P＜0.01).RT-PCR检测结果显示:不同浓度H2 O2处理SRA01/04细胞后,细胞中的miR-34a表达水平呈剂量依赖性升高,而SIRT1表达水平呈相应下降,与0μmol·L-1 H2O2组相比差异均有统计学意义(均为P ＜0.001).结论 在一定浓度H2O2诱导的人晶状体上皮细胞氧化应激中,miR-34a表达水平显著增加,而SIRT1表达水平显著下降.下调miR-34a表达可增加氧化应激人晶状体上皮细胞存活率.

Expression of miR-34a/SIRT1 in human lens epithelial cells during H2O2-induced oxidative stress

Objective To investigate the expression of microRNA-34a (miR-34a) and silent information regulator 1 (SIRT1) in human lens epithelial cells under H2O2-induced oxidative stress.Methods Different concentrations of H2O2 (0 μmol · L-1,100 μ mol· L-1,200 μmol · L-1,300 μmol · L-1,and 400 μmol · L-1) were used to stimulate SRA01/04 cells for 24 hours.Cell viability was measured using cell counting kit-8 (CCK-8) assay.Cell apoptosis was detected by flow cytometry.Expression levels of miR-34a/SIRT1 were measured by RT-PCR.Results CCK-8 assay showed that a certain concentration range of H2O2 had a proliferation inhibition on SRA01/04 cells.There was a dose response relationship between 100 μmol · L-1 and 400 μmol · L-1.Compared with 0 μmol · L-1 H2O2 group,the difference was statistically significant (all P ＜ 0.01).According to flow cytometry results,apoptotic rate of SRA01/04 cells in control group and H2O2(100-300 μmol · L-1) groups were (6.1 ± 1.2)％,(26.3 ± 1.8)％,(32.5 ± 2.2) ％,and (64.7 ± 5.3) ％.Compared with 0 μmol · L-1 H2 O2 group,the differences were statistically significant (all P ＜ 0.01).RT-PCR test results showed that the expression of miR-34a increased significantly in a dose-dependent manner after the SRA01/04 cells treated with different concentrations of H2O2,while SIRT1 expression level was decreased,there were significant differences compared with control group (all P ＜ 0.001).Conclusion There is a significantly increase of miR-34a and decrease of SIRT1 in human lens epithelial cells under the oxidative stress of a certain concentration of H2O2.Down-regulated expression of miR-34a can increase the survival rate of human lens epithelial cells under H2O2-induced oxidative stress.