绿茶多酚主要成分对百草枯诱导PC12细胞损伤的保护作用  

PROTECTIVE EFFECTS OF EGCG ON PARAQUAT-INDUCED INJURY IN PC12 CELLS

作　　者：厚荣荣[1] 陈建宗[1] 陈宏[1] 康小刚[1] 李源丽[1] 张颖[1] 李海龙[1] 

机构地区：[1]第四军医大学西京医院中医药研究中心,中西医结合老年脑病研究室,西安710033

出　　处：《神经解剖学杂志》2007年第1期83-87,共5页Chinese Journal of Neuroanatomy

摘　　要：探讨绿茶多酚的主要成分表没食子儿茶素没食子酸酯[(-)-epigallocatechin-3-gallate,EGCG]对百草枯(paraquat,PQ)诱导的PC12细胞损伤的影响及可能机制。建立PQ损伤的PC12细胞体外模型,经不同浓度EGCG预处理后,分别用四甲基偶氮唑盐(MTT)比色法检测细胞活性,Hochest33258染色观察凋亡时细胞核形态的变化,流式细胞仪(FCM)检测细胞凋亡比例,免疫细胞化学染色观察细胞色素c蛋白表达水平。1、5、10μmol/LEGCG对PQ诱导的PC12细胞损伤具有一定的保护作用。与PQ组(54.6±3.5%)相比,1、5、10μmol/LEGCG预处理组细胞活力分别上升为66.5±2.7%、74.4±2.6%、83.7±3.2%。Ho-chest33258染色发现PQ组较多胞核出现固缩凝集(43.5±8.4%),而EGCG预处理组则较少(分别为30.7±7.9%、17.9±6.8%、11.2±4.4%)。FCM检测也提示EGCG预处理可降低细胞的凋亡百分率,正常组、PQ组、EGCG预处理组(1、5、10μmol/L)分别为4%、39.9%、32.6%、20.1%和10.4%。另外,免疫细胞化学染色发现EGCG预处理可下调PQ诱导的细胞色素c在胞浆中的过表达。以上结果提示EGCG可减轻PQ诱导的PC12细胞损伤,其机制可能与下调细胞色素c在胞浆内的过表达有关。The present study was to observe the protective effects and related mechanisms of （-） - epigallocatechin-3-gallate （EGCG） a- gainst the PC12 cells injury induced by paraquat （PQ）. We established an in vitro model induced by PQ in PC12 cells. After pre-treatment with different concentration of EGCG, MTI＂ assay was used to detect the cell viability, Hcechst 33258 staining was employed to observe morphological changes of the cell nuclear, FCM was used to detect the apeptosis ratio, and immunocytochemical staining was adopted to observe the expression levels of cytochrome c. 1, 5, 10 μmol/L EGCG respectively played a protective role against PQ-induced PC12 cells injury. Compared with PQ-treated cells （ 54.6 ± 3.5 % ）, after exposure to pretreat groups with 1,5, 10 μmol/L EGCG, cell viability increased to 66.5 ±2.7%, 74.4 ±2.6%, 83.7 ±3.2%, respectively. Hochest 33258 staining demonstrated that more cells were characterized by nuclear condensation in PQ group（43.5 ± 8.4% ）, while in the group of EGCG, the rate was decreased to 30.7 ± 7.9%, 17.9 ± 6.8% and 11.2 ± 4.4%. FCM analysis indicated that apeptosis ratio was reduced by EGCG pretreatment, which in control, PQ and EGCG group （ 1,5,10 μ mol/L） was 4%, 39.9% and 32.6%, 20.1%, 10.4%, respectively. Besides, immunecytochemical staining indicated that EGCG downregulated the over-expression of cytochrome c in the cytosol induced by PQ. The data indicate that EGCG has a protective effect on the injury of PC12 cells induced by PQ, which may be related to downregulating the over-expression of cytochrome c in the cytosol.　

关 键 词：EGCG PC12细胞 百草枯 凋亡 神经保护 

分 类 号：R285[医药卫生—中药学]