LncRNA NALT通过调控Notch信号通路对H2O2诱导的人晶状体上皮细胞损伤的影响

Effects of lnc RNA NALT on H2O2-induced injury of human lens epithelial cells by regulating Notch signaling pathway

目的探讨lncRNA NALT(简称NALT)对H_(2)O_(2)诱导的人晶状体上皮细胞损伤的影响及其作用机制。方法使用不同浓度H_(2)O_(2)处理人晶状体上皮SRA01/04细胞24 h,CCK-8和qRT-PCR检测细胞增殖活性及NALT表达水平。将NALT过表达质粒转染至SRA01/04细胞中,再经100μmol/L H_(2)O_(2)或联合10μmol/L Notch信号通路抑制剂DAPT干预24 h,CCK-8检测细胞增殖活性;DCFH-DA荧光探针标记法检测各组细胞ROS水平;化学法检测SOD活性和MDA含量;Annexin V-FITC/PI法检测细胞凋亡情况;Western blot检测细胞中Notch1和Hes1蛋白表达水平。结果随着H_(2)O_(2)干预浓度的增加,SRA01/04细胞增殖活性和NALT表达水平均呈剂量依赖性降低(P<0.05)。H_(2)O_(2)干预后,NALT过表达可显著提高SRA01/04细胞增殖活性和SOD活性,降低ROS、MDA及细胞凋亡水平,上调Notch1和Hes1蛋白表达水平(均P<0.05)。然而,联合DAPT干预可逆转NALT过表达对H_(2)O_(2)诱导的SRA01/04细胞损伤的保护作用。结论过表达NALT可改善H_(2)O_(2)诱导的人晶状体上皮细胞损伤,其机制可能与激活Notch信号通路有关。

Objective The aim of this study was to investigate the effects and its mechanism of lncRNA NALT(NALT)on the injury of human lens epithelial cells induced by H_(2)O_(2).Methods Human lens epithelial SRA01/04 cells were treated with different concentrations of H_(2)O_(2)for 24 h,and the cell proliferation activity and NALT expression level were detected by CCK-8 and q RT-PCR.The NALT overexpression plasmid was transfected into SRA01/04 cells,and then treated with 100μmol/L H_(2)O_(2)or combined with 10μmol/L Notch signaling pathway inhibitor DAPT for 24 h.The cell proliferation activity was detected by CCK-8.The level of ROS was evaluated by DCFH-DA fluorescent probe labeling method.The SOD activity and MDA content were assessed by chemical method.The level of cell apoptosis was detected by Annexin V-FITC/PI method.The protein expression levels of Notch1 and Hes1 were evaluated by Western blot.Results With the increase of H_(2)O_(2)concentrations,the proliferation activity and NALT expression level of SRA01/04 cells were decreased in a dose-dependent manner(P<0.05).After H_(2)O_(2)intervention,NALT overexpression increased the proliferation activity and SOD activity of SRA01/04cells,decreased the level of ROS,MDA and apoptosis,and up-regulated the protein expression levels of Notch1 and Hes1(P<0.05).However,combined DAPT intervention reversed the protective effects of NALT overexpression on SRA01/04 cell injury induced by H_(2)O_(2).Conclusion Overexpression of NALT improved the injury of human lens epithelial cells induced by H_(2)O_(2),and the mechanism may be related to the activation of Notch signaling pathway.