NADH对X射线照射正常肝细胞L02后PARP和Caspase 3活性的影响

摘要

Objective To study the influence of NADH on cell apoptosis induced by X-ray and the activity of PARP and Caspase 3. Methods This experiment established the pseudo-irradiation group, control group and treatment group.RPMI 1640 medium with and without NADH(400μg/mL)was respectively added to the normal hepatocyte of the control group and treatment group after the normal hepa-tocyte was irradiated under 6MV varian 5.0Gy.The normal hepatocyte was cultured for 24hours.The apoptosis rate was tested by Annexin V/PI, bcl-2 and Bax protein positive expression rate were tested by flow cytometry, the activity of poly-ADP-ribose polymerase PARP and Caspase3 were tested by western blot.Results Comparing with the control group, the apoptosis rate obviously reduced, Bax protein positive expression percentage apparently reduced but the bcl-2 expression percentage obviously increased with ( P<0.05 ) .Both the PARP and Caspase 3 of the control group did not emerge cut fragment and the treatment group did not find the cut fragment.Conclusion NADH can resist the damage and apoptosis of L02 normal hepatocyte reduced by the radiation.Its mode of action may be associated with the mitochondria of bcl-2 and bax.%目的：为了研究NADH对X射线诱导的细胞凋亡和PARP、Caspase 3活性的影响。方法设假照射组、对照组及处理组。处理组正常肝细胞在6MV Varian 5．0Gy照射后立即加入含NADH（400μg／mL） RPMI 1640完全培养基，假照射组和对照组只加入培养液，培养24 h，用Annexin V／PI法检测细胞凋亡率；流式细胞仪检测bcl－2和bax蛋白阳性表达率；免疫印迹检测PARP和Caspase3活性。结果处理组与对照组相比细胞凋亡率明显下降， bax蛋白阳性表达百分率明显下调，而bcl－2表达则明显上调（P＜0．05）。对照组PARP和Caspase 3蛋白均出现剪切片断，处理组未发现剪切片断形成。结论 NADH具有抗辐射诱导的L02肝细胞凋亡损伤作用，其作用机制可能通过bcl－2和bax调控线粒体途径有关。