摘要
目的探讨儿茶素没食子酸酯(EGCG)对过氧化氢(H_2O_2)诱导人视网膜色素上皮(ARPE-19)细胞氧化应激损伤及TRAF6/TAK1信号通路活化的影响。方法采用H_2O_2(100μmol/L)体外诱导ARPE-19细胞氧化应激损伤模型,将ARPE-19细胞分为正常对照组、H_2O_2诱导组、H_2O_2+低剂量(40μmol/L)EGCG组、H_2O_2+中剂量(80μmol/L)EGCG组、H_2O_2+高剂量(160μmol/L)EGCG组;CCK8法检测ARPE-19细胞相对存活率,相关试剂盒检测氧化应激产物MDA、NO水平及SOD1活性,Western Blot方法分析HO-1、SOD1、TRAF6、TAK1及p-TAK1蛋白表达。结果与H_2O_2诱导组比较,不同浓度EGCG干预的ARPE-19细胞相对存活率明显升高(P<0.05),细胞匀浆氧化应激产物MDA、NO水平明显降低(P<0.05),匀浆中SOD1活性显著升高(P<0.05),细胞中HO-1及SOD1表达水平显著升高(P<0.05),细胞中TRAF6及p-TAK1蛋白表达显著降低(P<0.05),并表现出剂量依赖性。结论 EGCG能明显抑制H_2O_2诱导的ARPE-19细胞氧化应激损伤,其作用机制可能与抑制TRAF6/TAK1信号通路活化有关。
OBJECTIVE To investigate the inhibitory effects of epigallocatechin gallate(EGCG) on oxidative stress injury induced by H2O2 in human retinal pigment epithelial(ARPE-19)cells and the activation of TRAF6/TAK1 signaling pathway. METHODS The H2O2(100 μmol/L)was used to induce the model of oxidative stress injury in ARPE-19 cells in vitro. All ARPE-19 cells were divided into five groups, including control group, H2O2-induced group, H2O2+low dose(40 μmol/L) EGCG group, H2O2+medium dose(80 μmol/L) EGCG group, and H2O2+high dose(160μmol/L) EGCG group. And CCK8 assay was used to measure the relative survival rates of ARPE-19 cells. Related kits were used to detect the levels of MDA, NO and the activity of SOD1. Western Blot was used to determine the expression levels of HO-1, SOD1, TRAF6, TAK1 and p-TAK1 proteins. RESULTS Compared with H2O2 induced group, the relative survival rates of ARPE-19 cells were greatly increased after treatment with different concentrations of EGCG(P<0.05). The levels of oxidative stress productions(MDA and NO) were significantly decreased, while the activity of SOD1 was greatly increased(P<0.05). The expression levels of HO-1 and SOD1 were also greatly increased, while the expression levels of TRAF6 and p-TAK1 were greatly decreased(P <0.05) with a dose-dependent manner.CONCLUSIONS The EGCG significantly inhibited the oxidative stress injury of ARPE-19 cells induced by H2O2 via suppressing the activation of TRAF6/TAK1 signaling pathway.
作者
刘艳
张懿
雷锐
周海艳
LIU Yan;ZHANG Yi;LEI Rui(The Central Hospital of Enshi Tujia and Miao Autonomous Prefecture, Enshi 445000, China)
出处
《中国中医眼科杂志》
2019年第2期107-111,共5页
China Journal of Chinese Ophthalmology
关键词
儿茶素没食子酸酯
人视网膜色素上皮细胞
氧化应激
信号通路
epigallocatechin gallate
human retinal pigment epithelial cells
oxidative stress
signaling pathway