ERCC6对过氧化氢诱导的晶状体上皮细胞线粒体质量调控的影响OA

Objective To investigate the effect of excision repair cross-complementing group 6(ERCC6)on mito-chondrial quality control in H2O2-induced human lens epithelial cells.Methods Human lens epithelial cells(SRA01/04 cells)were divided into the NC group,H2O2 group,H2O2+empty vector group,and H2O2+ERCC6 group.Except for the NC group,all other groups were treated with 400 μmol·L-1 H2O2.The latter two groups were transfected with blank plasmids and ERCC6 overexpression plasmids,respectively.Western blot was used to detect the expression of Cockayne syndrome B protein(CSB),mitochondrial transcription factor A(TFAM),dynamin-related protein 1(DRP1),mitofusin 1(MFN1),PINK1,Parkin,and LC3B proteins;RT-PCR was performed to determine mitochondrial DNA(mtDNA)copy number and the mRNA levels of DRP1 and MFN1;and immunofluorescence was used to measure mitochondrial membrane potential and mi-tochondrial reactive oxygen species(mtROS)levels.Results Compared with the NC group,the H2O2group showed de-creased CSB,TFAM and MFN1 protein expression,mtDNA copy number,MFN1 mRNA,and mitochondrial membrane poten-tial,whereas the relative expression levels of DRP1 mRNA and protein,mtROS,PINK1,Parkin,and LC3B protein increased(all P＜0.05).No significant differences were observed in the above indicators between the H2O2 group and the H2O2+empty vector group(all P＞0.05).Compared with the H2O2 group,the H2O2+ERCC6 group exhibited increased CSB,TFAM and MFN1 protein expression,mtDNA copy number,MFN1 mRNA,and mitochondrial membrane potential,whereas the rela-tive expression levels of DRP1 mRNA and protein,mtROS,PINK1,Parkin,and LC3B protein decreased(all P＜0.05).Con-clusion ERCC6 overexpression can promote mitochondrial biogenesis,correct mitochondrial dynamic imbalance,reduce mtROS,restore mitochondrial membrane potential,and inhibit PINK1/Parkin-mediated mitophagy,thereby improving mito-chondrial quality control in lens epithelial cells under oxidative stress.