表没食子儿茶素没食子酸酯对丙烯酰胺致小脑颗粒神经元凋亡的影响  

Protective effect of epigallocatechin-3-gallate on apoptosis of rat cerebellar granule neurons induced by acrylamide

作　　者：刘春芳[1] 江城梅[2] 周礼华[2] 

机构地区：[1]蚌埠医学院第一附属医院,安徽蚌埠233004 [2]蚌埠医学院预防医学系,安徽蚌埠233030

出　　处：《中南大学学报（医学版）》2012年第9期944-950,共7页Journal of Central South University ：Medical Science

基　　金：安徽省教育厅自然科学研究项目(2006kj345B)~~

摘　　要：目的:探讨表没食子儿茶素没食子酸酯(EGCG)对丙烯酰胺(ACR)致小脑颗粒神经元(CGNs)凋亡的保护作用。方法:体外培养的CGNs经不同浓度EGCG(5,10,25,50,100μmol/L)预处理48h后,加入ACR(5mmol/L)染毒24 h,用四甲基偶氮唑盐(M)比色法检测细胞存活率,测定细胞内超氧化物歧化酶(SOD)活性及丙二醛(MDA)的含量,Hochst33342荧光显微镜染色观察凋亡时细胞核形态的变化,RT-PCR检测细胞bcl-2 mRNA与bax mRNA表达情况。结果:同ACR损伤组比较,终浓度为10,25,50μmol/L的EGCG能减轻ACR引起的CGNs的损伤,明显提高细胞的存活率(P<0.05),SOD活性增高,MDA含量降低。Hoechst33342染色发现细胞核固缩、致密浓染现象较ACR损伤组改善明显,bcl-2mRNA表达增强,bax mRNA表达减弱,bcl-2/bax比值降低(P<0.05),25μmol/LEGCG组保护效果最佳(P<0.01),而终浓度为100μmol/LEGCG组无明显保护作用。结论:EGCG在一定剂量范围内对ACR引起的CGNs凋亡有保护作用。Objective： To investigate the protective effect of epigallocatechin-3 -gaUate （EGCG） on apoptosis ofcerebellar granule neurons （CGNs） induced by acrylamide （ACR）. Methods： CGNs were cultured with the addition of 5 mmol/L ACR for 24 hours to set up a cell injury model. Prior to ACR treatment, CGNs were treated with different concentrations of EGCG （0, 5, 10, 25, 50, 100 [xmol/L） for 48 hours. Neuronal viability was measured with metylthiazdyltetrazolium （MTT）. The activity of SOD and the content of MDA were assayed. Hoechst33342 staining was employed to observe morphological changes of the cell nucleus. Reverse transcription-polymerase chain reaction （RT-PCR） was used to measure expression ofbcl- 2 mRNA and bax mRNA.Results： At the concentrations of 10, 25 or 50 μmol/L, EGCG played a protective role against ACR- induced CGN injury. Compared with ACR injured group （no EGCG）, EGCG improved the cell , enhanced SOD activit decreased the level of MDA as well as the cell apoptosis ratio （P〈0.05）. Bd-2 mRNA expression was increased and bax mRNA expression was reduced （P〈0.05）. 25 μmol/L EGCG had the largest effect. However, 100 μmol/L EGCG did not have a significantly protective effect. Conclusion： EGCG at appropriate concentration has protective effect against the CGNs on apoptosis induced by ACR.

关 键 词：表没食子儿茶素没食子酸酯 丙烯酰胺 小脑颗粒神经元 细胞凋亡 神经保护 

分 类 号：R285[医药卫生—中药学]