Exendin-4对t-BHP诱导的SV40细胞氧化应激的影响

The Effect of Exendin-4 on the Oxidative Stress of SV40 Cells Induced by t-BHP

目的探讨肠促胰岛素类似物Exendin-4对t-BHP诱导下小鼠肾小球系膜细胞氧化应激的影响。方法培养小鼠肾小球系膜细胞SV40,并进行分组:(1)对照组,予含有5%FBS、糖浓度5.5mmol/L的DMED培液培养;(2)50及100μmol/L t-BHP干预组(T50及T100组),分别予50或100μmol/L的t-BHP干预4h;(3)Exendin-4预处理组(ET50及ET100组),50nmol/L Exendin-4预处理2h后,再予50或100μmol/L的t-BHP干预4h。采用CCK-8法检测细胞存活率,WST-8法测定细胞内超氧化物歧化酶(SOD)活性,分光光度法检测细胞内Caspase-3及谷胱甘肽还原酶(GR)活性,Western-blot测定Bax及Bcl-2的表达。结果 t-BHP处理后,细胞存活率及细胞内SOD及GR活性降低,抗凋亡基因Bcl-2表达下降,促凋亡基因Bax表达增加,Caspase-3活性升高。加用Exendin-4预处理后,细胞存活率及细胞内SOD,GR活性以及Bcl-2表达均较t-BHP单独处理组升高,Caspase-3活性及Bax表达均较t-BHP单独处理组降低,与对照组比较差别无统计学意义。结论 Exendin-4能改善t-BHP诱导的氧化应激,对SV40细胞具有保护作用。

Objective To investigate the effect of Exendin-4 on the oxidative stress of glomerular mesangial cells induced by t-BHP in mice. Methods Cultured mouse glomerular mesangial cells SV40 were divided into control group,50 and 100μmol/L t-BHP alone group,and 50 nmol/L Exendin-4 pretreatment group,The cell viability was measured by CCK-8 method. The activity of SOD was measured by WST-8 method. The activity of Caspase-3 and glutathione reductase were detected by spectrophotometry. The expression of Bax and Bcl-2 was detected by Western-blot. Results The cell viability and the activity of SOD and glutathione reductase decreased,the expression of anti-apoptotic gene Bcl-2 decreased,the expression of Bax was increased,the activity of Caspase-3 increased. After the pre-treatment with Exendin-4,the activity of SOD and glutathione reductase and the expression of Bcl-2 were higher than those in t-BHP alone group,while the expression of Caspase-3 and Bax were lower than those in the control group. The changes compared to the control group were not significant statistically. Conclusion Exendin-4 can improve t-BHP-induced oxidative stress and protect SV40 cells.