PARP-1蛋白在镉致NRK-52E细胞凋亡中的作用研究

Effects of PARP-1 protein overexpression on apoptosis induced by cadmium in NRK-52E cells

为了探究PARP-1蛋白在镉(Cd)致大鼠肾小管上皮细胞(NRK-52E)凋亡中的作用。本研究用RTCA技术检测Cd对细胞增殖的影响,用Western Blot方法检测细胞中PARP-1与AIF蛋白的表达情况,用流式细胞术或免疫荧光的方法检测Cd与NAC或DPQ共孵育对细胞凋亡率、线粒体膜电位以及活性氧的影响。结果显示,Cd影响了细胞的增殖并上调PARP-1蛋白的表达和活性氧(ROS)的生成,抑制PARP-1的活性后细胞凋亡率下降并且线粒体膜电位与细胞内ROS的含量都恢复到相对正常水平;添加NAC处理后减弱了Cd对细胞增殖的抑制作用同时降低了细胞凋亡率。结果表明,PARP-1蛋白和氧化应激在Cd致NRK-52E细胞凋亡中发挥了重要作用。

In this study, we investigated the effects of PARP - I on cadmium （Cd） - induced apoptosis of rat renal tubular epi-thelial cells （ NRK -52E）. The effect of Cd on cell proliferation was measured by RTCA technique; the expression of PARP - 1 andAIF protein as analyzed by Western Blot ; at the same time, the effects of co - incubation of Cd with NAC or DPQ on apoptosis rate,mitochondrial membrane potential and reactive oxygen species （ROS） were examined by flow cytometry or immunofluorescence. TheResults showed that Cd affected cell proliferation and promotes PARP - 1 protein expression and ROS production; apoptosis rateswere reduced after inhibiting the activity of PARP - 1 protein, and mitochondrial membrane potential and intracellular Ros contentreturned to a relatively normal level ; addition of NAC treatment reduced the inhibitory effect of Cd on cell proliferation and decreasedthe apoptotic rate. It could concluded that the PARP - 1 protein and oxidative stress play an important role in Cd - induced NRK -52E cell apoptosis.