白藜芦醇对曲格列酮诱发HepaRG细胞氧化损伤的影响

摘要

目的：探讨白藜芦醇对曲格列酮诱发HepaRG细胞氧化损伤的影响及可能作用机制。方法实验分组包括：正常对照组（含有0．1％DMSO的RPMI 1640培养基）、50μmol／L曲格列酮组、白藜芦醇3种浓度（3．75，7．5，15μmol／L）与50μmol／L曲格列酮的共处理组。 MTT法检测曲格列酮、白藜芦醇以及白藜芦醇与50μmol／L曲格列酮共同作用对HepaRG细胞的增殖抑制作用。以上各组作用48 h后检测各组细胞内活性氧（ reactive oxygen spe－cies， ROS）的含量，脂质过氧化（ lipid peroxidation ）和细胞凋亡程度，细胞的总抗氧化能力，以及细胞内过氧化氢酶（catalase， CAT）、谷胱甘肽过氧化物酶（glutathione peroxidase， GSH－px）、超氧化物歧化酶（superoxide dismutase， SOD）的活性。结果曲格列酮能明显导致HepaRG细胞产生氧化应激现象。与正常对照组相比，曲格列酮处理组ROS和脂质过氧化产物丙二醛（malondialdehyde，MDA）水平以及细胞凋亡和坏死率均显著升高（P＜0．05），总抗氧化能力大幅降低（P＜0．05），CAT、GSH－px、SOD的活性均降低（P＜0．05）。加入不同浓度白藜芦醇共同作用后， ROS和MDA生成量有所下降（P＜0．05），细胞凋亡和坏死率也相应下降（P＜0．05），细胞总抗氧化能力以及以上3种抗氧化物酶的活性均有所提高（P＜0．05），且有一定的剂量依赖关系。结论曲格列酮能引起HepaRG细胞产生明显的氧化应激作用，白藜芦醇能够显著改善由曲格列酮对 HepaRG细胞所造成的氧化损伤。%Objective To explore the effect of resveratrol ( Rev) as an antioxidant on oxidative damage to HepaRG cells induced by troglitazone ( Tro).Methods Cells were divided into five groups: control ( RPMI 1640 only with 0.1%DMSO), Tro(50 μmol/L), Tro(50 μmol/L) +Rev(15 μmol/L), Tro(50 μmol/L) +Rev(7.5 μmol/L) and Tro (50 μmol/L)+Rev(3.75 μmol/L) groups.MTT assay was performed to detect the viability of Rev-treated, Tro-treated and Rev with 50 μmol/L Tro-treated HepaRG cells.After 48 hours, the level of reactive oxygen species (ROS) and lipid oxidation ( malondialdehyde , MDA ) , degree of apoptosis , total antioxidant capacity , activity of hydrogenperoxidase (catalase, CAT), glutathione peroxidase (GSH-px) and superoxide dismutase(SOD)of these groups were identified. Results Tro could obviously cause HepaRG cells to produce oxidative stress .Compared with control group ,ROS and lipid peroxidation ( MDA) levels and the rate of apoptosis and necrosis in Tro-treated group were significantly increased ( P<0.05),total antioxidant capacity greatly reduced (P<0.05),and the activity of CAT,GSH-px and SOD was decreased (P<0.05).After adding various concentrations of Rev interaction , ROS and MDA production volume decreased (P < 0.05), and the apoptosis and necrosis rate correspondingly declined (P<0.05).Total antioxidant capacity of the cells and the activity of the three antioxidant enzymes were increased (P<0.05), and there was a dose-dependent relationship. Conclusion Tro can cause HepaRG cells to produce significant oxidative stress while Rev can significantly improve the oxidative damage of Tro to HepaRG cells .