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目的探索祁州漏芦乙醇提取物(RUEE)对过氧化氢(H2O2)致肝细胞损伤的保护作用。方法以H2O2损伤Chang Liver细胞建立氧化应激模型,以不同浓度RUEE(12.5~400 mg/L)进行干预。采用MTT法检测细胞存活率;以微板比色法检测培养液中LDH和转氨酶活性以及细胞中还原型谷胱甘肽(GSH)和丙二醛(MDA)含量;免疫印迹技术检测核因子-E2相关因子2(Nrf2)蛋白含量。结果 RUEE在12.5~100 mg/L范围内对Chang Liver细胞存活率无显著影响,不具细胞毒作用。与对照组相比,H2O2组细胞内氧化应激增强,细胞损伤显著。RUEE能剂量依赖性地提高细胞存活率,降低细胞培养液中LDH、ALT、AST的释放;降低细胞MDA含量,升高细胞GSH含量,增高核中Nrf2蛋白水平。结论祁州漏芦对H2O2致肝细胞氧化应激损伤具有保护作用。
Objective To explore the protective effect of RUEE on liver cell injury induced by hydrogen peroxide (H2O2). Methods Oxidative stress was induced by H2O2 in Chang Liver cells. Interventions were given at different concentrations of RUEE (12.5-400 mg / L). Cell viability was detected by MTT assay. LDH and aminotransferase activities, glutathione (GSH) and malondialdehyde (MDA) contents in culture medium were detected by microplate colorimetric assay. Nuclear factor- E2 related factor 2 (Nrf2) protein content. Results RUEE had no significant effect on the viability of Chang liver cells in the range of 12.5 ~ 100 mg / L, with no cytotoxic effect. Compared with the control group, H2O2 group intracellular oxidative stress increased significantly cell damage. RUEE could increase cell viability in a dose-dependent manner, decrease the release of LDH, ALT and AST in cell culture medium, decrease the content of MDA, increase the content of GSH and increase the level of Nrf2 in the nucleus. Conclusion Leizhou gourd can protect liver cells from oxidative stress induced by H2O2.