槲皮素对大鼠原代肝细胞酒精性氧化损伤的防护作用

目的:观察槲皮素(quercetin)对大鼠原代肝细胞酒精性氧化损伤的防护作用。方法:经二步胶原酶技术分离培养大鼠原代肝细胞,用100mmol/L无水乙醇染毒细胞8h,作为酒精损伤模型。乙醇染毒前经不同剂量槲皮素(25～200μmol/L)预作用不同时间(0～8h),观察槲皮素干预对细胞培养上清液中乳酸脱氢酶(LDH)、谷草转氨酶(AST)活性,丙二醛(MDA)、谷胱甘肽(GSH)水平和超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性的影响。结果:大鼠原代肝细胞经100mmol/L酒精暴露8h后,LDH、AST活性,MDA、GSH含量和SOD、CAT活性与对照组相比,均有非常显著性差异;酒精暴露前1～4h经50～75μmol/L槲皮素干预效果最明显,LDH、AST、MDA和GSH、SOD、CAT水平分别较酒精组非常显著性降低和升高。结论:槲皮素可能通过减少GSH耗竭,提高抗氧化酶活性,抑制脂质过氧化来保护大鼠原代肝细胞抗酒精性氧化损伤,保护作用呈剂量与时间效应关系。

PROTECTION OF QUERCETIN AGAINST ETHANOL-INDUCED OXIDATIVE DAMAGE IN RAT PRIMARY HEPATOCYTES

Objective:To investigate the protective effects of quercetin on ethanol-induced oxidative damage in rat primary hepatocytes. Method:Rat primary hepatocytes were isolated and cultured by using two steps collagenase perfusion technique. The hepatocytes were pre-treated with quercetin of different doses (25-200 μmol/L) in different time (0-8 h) followed by sensitive 100 mmol/L ethanol exposure for 8 h. Then LDH, AST activities in the supernatants of hepatocytes, and MDA, GSH contents, SOD, CAT activities in hepatocytes were determined. Results:There were significant changes of AST, LDH, MDA, GSH, SOD and CAT between ethanol group and normal control group. However, the pre-treatment of quercetin at the dose of 50-75 μmol/L for 1-4 h could significantly prohibit the release of AST and LDH, the rise of MDA level and the decline of SOD, CAT activities and GSH content of hepatocytes exposed to ethanol. Conclusion:Quercetin could prevent rat primary hepatocytes from ethanol-induced oxidative damage probably by reducing GSH consumption, improving antioxidant enzyme activity and inhibiting lipid peroxidation reaction at dose-dependent and time-dependent manner.