表没食子儿茶素没食子酸酯对心肌细胞缺氧/复氧损伤的抑制作用及其机制  

Protective effect of epigallocatechin-3-gallate on hypoxia/reoxygenation injury in cardiac myocytes and its mechanism

作　　者：郑晓佳 余婷 张华[1] 李春凤[2] 陈叶霞 阳勇珍 李晨昕 简洁[1] 

机构地区：[1]桂林医学院药学院,广西桂林541004 [2]桂林医学院附属医院感控科,广西桂林541004

出　　处：《中国药理学通报》2017年第11期1584-1588,共5页Chinese Pharmacological Bulletin

基　　金：广西高校大学生创新创业计划项目(No 201510601007);广西高等学校优秀人才资助计划项目

摘　　要：目的探讨表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate,EGCG)对H9c2心肌细胞缺氧/复氧损伤的抑制作用及其机制。方法体外培养H9c2心肌细胞,随机分为正常(N)、缺氧/复氧(H/R)、EGCG低剂量(L)、中剂量(M)和高剂量组(H),共5组(n=6)。建立细胞缺氧/复氧模型,给予EGCG预处理,用CCK-8法测定细胞存活率;Annex V-FITC/PI双染法检测细胞凋亡率;按照试剂盒说明书检测细胞培养液T-AOC、TNF-α含量;用Western blot法检测Akt蛋白及磷酸化水平;用荧光定量PCR法检测PI3K、Akt、caspase-3基因表达。结果与模型组相比,EGCG能增加H9c2心肌细胞缺氧/复氧损伤后的存活率,减少细胞凋亡,提高T-AOC水平,降低TNF-α含量,增加PI3K、Akt和p-Akt表达,减少caspase-3的表达。结论 EGCG通过影响PI3K/Akt信号通路,减少细胞凋亡,保护心肌细胞。Aim To observe the protective effect of epigallocatechin-3-gallate( EGCG) on hypoxia/reoxygenation( H/R) injury of cardiac myocytes and its mechanisms. Methods H9 c2 cardiac myocytes were cultured in vitro and randomly divided into five groups:normal group( N group),H/R group,EGCG low dose group( L group), EGCG medium dose group( M group),and EGCG high dose group( H group). The cardiomyocyte H/R injury model was established and EGCG was pretreated. Cell survival rate was tested by CCK-8 method. The cell apoptotic rate was detected using Annexin V-FITC/PI double staining. The contents of total antioxidant capacity( T-AOC) and tumor necrosis factor α( TNF-α) in cell culture medium were tested according to the kit instructions. The protein expression of Akt and p-Akt was observed using Western blot,while the gene expressions of PI3 K,Akt,caspase-3 were detected by using fluorescence quantitative PCR method. Results Compared with model group,EGCG increased cell survival rate and reduced the apoptosis after H/R injury. Meanwhile,pretreatment EGCG improved the activity of T-AOC,reduced the level of TNF-α,up-regulated the expression of PI3 K,Akt and p-Akt,and down-regulated the expression of caspase-3. Conclusion EGCG reduces apoptosis and protects cardiac myocytes by influencing PI3 K/Akt signal pathway.

关 键 词：EGCG H9C2心肌细胞 缺氧/复氧损伤 细胞凋亡 PI3K/Akt caspase-3 

分 类 号：R-332[医药卫生] R282.71