姜黄素对多巴胺诱导PC12细胞凋亡的拮抗作用

Protective effect of curcumin on dopamine-induced apoptosis in PC12 cells

目的探讨姜黄素（Cur）对多巴胺氧化应激损伤PC12细胞的保护作用。方法以多巴胺（DA）损伤PC12细胞为氧化应激损伤的模型,采用MTT法检测细胞增殖状况,Hoechst染色检测细胞核形态,碘化丙啶（PI）染色流式细胞术（FCM）检测细胞凋亡,罗丹明123（Rh123）染色FCM检测细胞线粒体膜电位（MMP）。结果多巴胺（50~800μmol/L,24h）呈浓度依赖性地损伤PC12细胞,姜黄素（10~40μmol/L）对PC12细胞无毒性作用,但呈浓度依赖性地减轻DA对PC12细胞的毒性作用,抑制DA诱导PC12细胞凋亡,减轻DA对PC12MMP的降低作用。结论姜黄素可抑制DA诱导PC12细胞凋亡,对DA氧化应激损伤神经元具有保护作用,其机制可能与姜黄素抑制MMP的降低有关。

Objective：To explore the protective effect of curcumin（Cur） on apoptosis of PC12 cells induced by dopamine（DA） oxidative stress.Methods：Using DA to induce PC12 cell damage,the model of oxidative stress-induced neuron damage was established.Proliferation of PC12 cells was observed by 3-[4,5-dimethylthiazolyl ]-2,5-diphenyltetrazolium bromide（MTT） assay.Morphology of the nucleus was observed by Hoechst staining.Apoptosis of PC12 cells was detected by propidium iodide（PI） staining flow cytometry（FCM）.The mitochondrial membrane potential（△Ψm） was determined by Rh123 staining flow cytometry.Results：① DA damaged PC12 cells in a concentration-dependent manner after application of DA（50-800 μmol/L） for 24 h,and MTT assay showed that Cur（10-40 μmol/L） had no toxic effect on PC12 cells.② Cur reduced the toxic effect induced by DA on PC12 cells in a concentration-dependent manner.③ Cur reducing the DA-induced toxic effect on PC12 cells was confirmed by cellular morphologic experiments.④ PI staining FCM showed that Cur inhibited cellular apoptosis induced by DA and this effect was also confirmed by Hoechst staining.⑤ The level of △Ψm was significantly decreased in PC12 cells after 100 and 200 μmol/L DA treatment for 24 h,which was significantly ameliorated by Cur（20 and 40 μmol/L） treatment.Conclusion：Cur has a protective effect on PC12 cell damage induced by DA oxidative stress and this effect may be ascribed to the amelioration of the decreasing level of △Ψm.