蛋白酶体β5过表达对人晶状体上皮细胞抗氧化能力的影响

The Effect of Overexpression of Proteasome PSMB5 on Antioxidative Ability of Human Lens Epithelial Cells

目的:研究蛋白酶体β5(PSMB5)过表达对人晶状体上皮细胞抗氧化能力的影响。方法:实验研究。构建重组质粒pcDNA3.1-PSMB5,将其转染入人晶状体上皮细胞株SRA01/04中,形成稳定表达(作为实验组),设pcDNA3.1空载体作为对照组。将2组细胞置于40 μmol/L H 2O 2环境下,Hoechst 33342荧光染色观察2组细胞核形态的变化;流式细胞仪检测2组细胞凋亡率的变化。数据采用独立样本 t检验。 结果:40 μmol/L H 2O 2环境下,Hoechst 33342荧光染色可见,与实验组相比,对照组的细胞核明显皱缩、变形;流式细胞仪检测发现,实验组和对照组的细胞凋亡指数分别为(9.3±0.9)%和(15.7±1.9)%,均高于处理前的(5.9±0.8)%和(7.1±1.1)%,对照组凋亡指数比实验组的凋亡指数更高( t=3.742, P=0.008)。 结论:PSMB5基因过表达能有效提高人晶状体上皮细胞的抗氧化能力。

Objective:To discuss the antioxidative ability of proteasome subunitβ5(PSMB5)over expression in human lens epithelial cells.Methods:This is an experimental study.Recombinant plasmid pcDNA3.1-PSMB5 was constructed and transfected into a human lens epithelium strain SRA01/04 to form a stable transfection(experimental group),and empty pcDNA3.1 was also transfected into lens epithelial cells at the same time as a control.Two groups of cells were treated with 40μmol/L H2O2.The nuclear morphology of the two group cells was observed by Hoechst 33342 stain,and cell apoptosis of the two group cells was dectected by flow cytometry.Data was analyzed by a paired t test.Results:Under a 40μmol/L H2O2 treated environment,Staining by Hoechst 33342 indicated that the nuclei in the control group were significantly shrunken and condensed compared with those of the experimental group.The apoptotic percentages detected by flow cytometry in the experimental group and control group were(9.3±0.9)%and(15.7±1.9)%,and both had increased compared to pre-treatment with H2O2,which were(5.9±0.8)%and(7.1±1.1)%,respectively.The apoptotic percentage in the control group increased to a higher level than in the experimental group(t=3.742,P=0.008).Conclusions:Overexpression of proteasome PSMB5 can effectively improve the antioxidative ability of human lens epithelial cells.