急性乙醇暴露对人原代肝细胞血红素氧化酶的影响

Effect of acute ethanol exposure on HO-1 enzyme activity in humen primary hepatocytes

目的 研究急性乙醇暴露对人原代肝细胞血红素氧化酶活力及蛋白水平的影响。方法 经体外灌流、分离培养人原代肝细胞 ,观察乙醇对人原代肝细胞上清液中天冬氨酸氨基转移酶 (AST)的释放及谷胱甘肽 (GSH)含量的变化 ,用westernblot方法检测乙醇对人原代肝细胞血红素氧化酶活力及蛋白水平的影响。结果 急性乙醇暴露导致人原代肝细胞上清液中释放的AST增加 ,并呈明显的剂量效应和时间效应关系 ;此外 ,在 10 0mmol L乙醇 2 4h暴露下 ,肝细胞中的GSH明显降低 ,而HO 1酶活力在 0 5～ 12h之间明显升高 ,随后开始降低 ,且HO 1蛋白水平变化趋势与此一致。结论 HO

Objective To study the effect of ethanol exposure on the expression of HO-1 enzymes in human primary culture hepatocytes. Methods Human primary hepatocytes were isolated from human liver tissue and cultured in Williams'E medium. After ethanol exposure, HO-1 enzyme activity and protein level were detected by spectrophotometry and western blotting, respectively. Meanwhile, the level of aspartate transaminase (AST) in the supernatants and GSH content in hepatocytes were measured to indicate cell oxidative-damage. Results There was remarkable dose- and time - dependent relationship between ethanol exposure and AST release, and a time -dependent decrease in GSH content. Moreover, we also observed that ethanol exposure could affect HO-1 enzyme activity and HO-1 protein level, at the beginning, it went up significantly then went down. Conclusion The increase of HO-1 enzyme activity and protein level after ethanol exposure may be relative to prevent against ethanol-induced oxidative injury in human primary hepatocytes.