没食子儿茶素没食子酸酯对MPP^＋诱导大鼠PC12细胞凋亡的拮抗作用

Antagonistic action of epigallocatechin-3-gallate on apoptosis in rat PC12 cells induced by 1-methyl-4-phenylpyridinium ion

目的探讨没食子儿茶素没食子酸酯(EGCG)对MPP+诱导大鼠PC12细胞凋亡的拮抗作用。方法培养PC12细胞给予MPP+(900μmol.L-1)诱导细胞凋亡,实验分为6组:空白对照组、MPP+组、维生素E(10μmol.L-1)组和EGCG高(100μmol.L-1)、中(50μmol.L-1)、低(10μmol.L-1)剂量组。药物处理0.5h后,加入MPP+损伤细胞,4h后取细胞测定乳酸脱氢酶(LDH)漏出量,MTT法检测细胞活力,Hoechst33342荧光染色法和流式细胞术检测细胞凋亡,透射电镜观察凋亡细胞线粒体形态结构改变。结果MPP+处理后,细胞活力下降,LDH漏出量增加,细胞凋亡率增加,线粒体肿胀,出现空泡和嵴断裂等细胞凋亡征象。维生素E和不同剂量EGCG处理后,明显提高细胞活力,降低LDH漏出和细胞凋亡率,并减少MPP+引起的线粒体结构损伤。结论EGCG具有抑制MPP+诱导的大鼠PC12细胞凋亡作用,其作用与保护细胞线粒体结构的完整性有关。

Aim To investigate the antagonistic action of EGCG on apoptosis of rat PC12 cell induced by MPP^＋. Methods PC12 cells were cultured and the apoptosis induced by MPP^＋ （900μmol · L^- 1 ） was ob- served. The cells were randomly divided into 6 groups ： blank group without any treatment, MPP^＋ control group, vitamin E group and EGCG groups（ 10,50,100 μmol · L^-1 ）. After treatment of drugs, cell v/ability,leakage of LDH, morphological changes of mitochondria and apoptosis were detected by MTT, Hoechst 33342 staining, transmission electron microscope and flow cytometry, respectively. Results After treatment of cultured PC12 cells with MPP^＋, cell viability was decreased, leakage of LDH and apoptotic rate were increased, and mitochondria swelling, vacuole and cris tae breakage were observed. Vitamin E and EGCG en-hanced cell viability, reduced the leakage of LDH and apoptotic rate, and decreased the damage degree of mitochondria. Conclusions EGCG possesses the ability of inhibiting rat PC12 cell apoptosis induced by MPP^＋ , and its protective action may relate to its function of keeping mitochondria integrality.