没食子儿茶素没食子酸酯对百草枯诱导SK-N-SH细胞凋亡的保护作用  

Protecting effect of epigallocatechin-3-gallate on apoptosis in SK-N-SH cells induced by paraquat

作　　者：李冰[1] 韩春明[2] 徐辰[3] 

机构地区：[1]北华大学基础医学院,吉林吉林132013 [2]暨南大学药学院,广东广州510632 [3]澳门理工学院高等卫生学校

出　　处：《中国新药与临床杂志》2010年第3期183-187,共5页Chinese Journal of New Drugs and Clinical Remedies

基　　金：澳门科技发展基金资助(035/2005/A)

摘　　要：目的探讨茶多酚中主要活性成分没食子儿茶素没食子酸酯(EGCG)对百草枯诱导人神经母细胞瘤细胞株SK-N-SH细胞凋亡的保护作用。方法培养的SK-N-SH细胞给予400μmol·L-1百草枯诱导细胞凋亡。实验分为6组:空白对照组、百草枯模型组、维生素E(10μmol·L-1)组和3个EGCG(1、5、10μmol·L-1)剂量组。以药物处理细胞2h后,加入百草枯,72h后MTT法检测细胞活力,取培养细胞上清液测定乳酸脱氢酶(LDH)漏出量,Hoechst 33258荧光染色法和流式细胞术检测细胞凋亡情况。结果与空白对照组相比,百草枯明显降低细胞活力(P<0.01),增加LDH漏出量(P<0.01),细胞膜结构不完整,出现空泡等凋亡现象,细胞凋亡发生率达到30.5%。EGCG处理后,显著提高细胞活力,减少LDH的漏出和降低细胞凋亡率(P<0.01),其中10μmol·L-1组的作用明显高于5μmol·L-1组或1μmol·L-1组(P<0.05或P<0.01)。结论EGCG具有抑制百草枯诱导的SK-N-SH细胞凋亡作用。AIM To investigate the protecting effect of epigallocatechin-3-gallate （EGCG） on apoptosis of human neuroblastoma （SK-N-SH） cells induced by paraquat. METHODS SK-N-SH cells were cultured and the cell apoptosis was induced by 400 μmol·L-1 paraquat. The cells were randomly divided into 6 groups： blank control group, paraquat model group, vitamin E （10 μmol·L-1） group and 3 EGCG （1, 5, and 10 μmol·L-1） groups. After treatment with Vitamin E and different concentration of EGCG for 2 hours respectively, 400 μmol. L -1 paraquat was added to the SK-N-SH cell cultured for 72 hours. Then MTT was used to determine the conditions of cells viability, and the supernatant was taken to determine the leakage of LDH. The morphological change and rate of cell apoptosis were detected by Hoechst 33258 staining and flow cytometry, respectively.RESULTS Compared with the blank control group, paraquat could reduce the cell viability （ P 0.01）,increase the leakage of LDH （P 0.01）, cause cell membrane breakage and vacuole, and induce apoptosis with a rate of occurrence up to 30.5%. With pre-treatment of EGCG, cell archaeus could be enhanced （P 0.01 or P 0.05）, the leakage of LDH was reduced （P 0.01）, and the rate of occurrence in cell apoptosis droped （P 0.01） significantly. And the effects of 10 μmol·L-1 EGCG group had significant difference with those of 5 μmol·L-1 group or 1 μmol·L-1 group （P 0.05 or P 0.01） . CONCLUSION EGCG can inhibit apoptosis of SK-N-SH cells induced by paraquat.

关 键 词：没食子儿茶素没食子酸酯 百草枯 细胞凋亡 SK-N-SH细胞 茶多酚 

分 类 号：R285.5[医药卫生—中药学]