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为探讨镉(Cd~(2+))致线粒体损伤在大鼠原代肾小管上皮细胞(r PT)凋亡中的作用,本研究对r PT细胞施以不同浓度梯度的Cd~(2+)经不同时间长短处理,并以添加PARP-1的抑制剂DPQ、抗氧化剂NAC处理作为对照处理,应用Western-blot的方法检测细胞中线粒体相关蛋白的表达,用免疫荧光的方法检测不同处理组中细胞凋亡率和线粒体膜电位的变化。结果显示,Cd~(2+)促进PARP-1蛋白的表达以及AIF和Cyt C从线粒体中释放;抑制PARP-1蛋白的活性后显著下调细胞凋亡率并且使线粒体膜电位恢复到相对正常水平;与Cd~(2+)组相比,Cd~(2+)+NAC处理组中相关促凋亡蛋白表达下降而抑凋亡蛋白表达增加,同时降低了细胞凋亡率,线粒体膜电位也恢复到相对正常水平。以上结果表明,Cd~(2+)使r PT细胞发生线粒体损伤以及氧化应激并且诱导细胞凋亡,其中PARP-1/AIF信号通路在这一过程中发挥了重要作用。
In order to investigate the role of cadmium (Cd 2+) -induced mitochondrial damage in rat primary renal tubular epithelial cells (r PT), r PT cells were treated with different concentration gradients of Cd 2+ ) Were treated with different lengths of time. PARP-1 inhibitor DPQ and antioxidant NAC were used as control. The expression of mitochondria-related protein in cells was detected by Western-blot and the expressions of mitochondria-associated protein In the rate of apoptosis and mitochondrial membrane potential changes. The results showed that Cd 2+ promoted the expression of PARP-1 protein and release of AIF and Cyt C from mitochondria. After inhibiting the activity of PARP-1 protein, the apoptosis rate was significantly decreased and the mitochondrial membrane potential returned to a relatively normal level ; Compared with Cd 2+ group, the expression of related pro-apoptotic protein decreased and the expression of pro-apoptotic protein increased in Cd 2+ (2 +) + NAC group, meanwhile decreased apoptosis rate, mitochondrial membrane potential Return to a relatively normal level. The above results indicate that Cd 2+ can induce mitochondrial damage, oxidative stress and induce apoptosis in r PT cells. PARP-1 / AIF signal pathway plays an important role in this process.