原花青素对过氧化氢损伤内皮细胞SOD、GSH-PX活性的影响

Effect of Procyanidins on the Activities of SOD,GSH -PX in Endothelial Cell Injured By Hydrogen Perdxide

目的观察原花青素对过氧化氢诱导人脐静脉内皮细胞(HUVEC)损伤的保护作用及其机制。方法体外培养人脐静脉内皮细胞,将细胞分为6组,即空白对照组、H2O2损伤组、H2O2+Vit C组、H2O2+原花青素5、25、50μmol/L组。将750μmol/L H2O2作用于加入Vit C及不同浓度原花青素预培养24 h的内皮细胞,继续培养18 h,采用四甲基偶氮唑盐比色法(MTT法)检测原花青素对H2O2诱导的内皮细胞氧化损伤的保护作用;用流式细胞仪分析-AnnexinV/PI双染色法检测细胞凋亡;定量检测各实验组超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)的活力。结果内皮细胞H2O2损伤后吸光度(OD)低于空白对照组,原花青素预处理后OD值增加且呈剂量依赖性;H2O2损伤可引起细胞SOD及GSH-PX活性明显降低,原花青素能维持SOD、GSH-PX活性,且呈剂量依赖性,各指标差异有显著性,并且呈剂量依赖性抑制H2O2诱导的内皮细胞的凋亡。结论原花青素能抑制过氧化氢诱导的血管内皮细胞的损伤及凋亡,其作用可能与维持SOD、GSH-PX活性有关。

Objective： To study the anti-injury effects of Procyanidins on the human umbilical vein endothelial celI（HUVEC） injured by hydrogen peroxide （H2O2） and its mechanism. Methods： Cultured human umbilical vein endothelial cells, the cells were divided into six groups：the control group, oxidative damage group, oxidative damage to join VitC control group,oxidative damage to add procyanidins 5 μmol/L,25 μmol/L,50 μmol/L concentration group. The 750 μmol/L H2O2 role Vit C and different concentrations of procyanidins 24 hours of pre-cultured endothelial ceils, cultured for 18 hours （cysteine protease expression determination time of 14 hours）, MTT assay was used to detect the protection of Procyanidins;to cell apoptosis was detected by Annexin V/PI flow cytometry, and the activities of each experimental group superoxodismutase （SOD）,glutathione peroxidase （GSH-PX） were quantitated. Results： The cell optical density in oxidation damaged group was lower than that in normal control. When pretreated by Procyanidins,the OD value was increased, and showed dose-dependent manner; H2O2 injury could cause cell SOD and GSH-PX activity significantly decreased;Procyanidins could maintain the activities of SOD, GSH-PX activity, and also showed dose-dependent manner;the various indicators of the difference was statistically significant, and Procyanidins could dose-dependently inhibit H2O2-induced apoptosis of endothelial cells. Conclusion： Procyanidins inhibit vascular endothelial cell apoptosis and injure induced by hydrogen peroxide and its role may be related to the maintaining of SOD, GSH-PX activities.