酒精对人原代培养肝细胞的氧化损伤与CYP 2E1关系的研究

目的 研究急性酒精暴露下对人原代培养肝细胞中CYP 2E1依赖的毒性作用和氧化损伤。方法 分离培养人原代肝细胞，以25-100mmol/L乙醇作用于人原代肝细胞9h及100mmol／L乙醇作用于人原代肝细胞0～24h后，检测人原代肝细胞中CYP 2E1的含量，并研100mmol/L乙醇作用于人原代肝细胞0～24h后，天冬氨酸转胺酶(aspartate transaminase，AST)的释放量及肝细胞中谷胱甘肽(Glutathione，GSH)、丙二醛(Malonclialdehyde，MDA)的含量。结果 急性酒精暴露导致人原代肝细胞中CYP 2E1的释放增加，并呈明显的剂量效应和时间效应关系；在100mmol/L乙醇作用下，AST和MDA明显升高，在0～24h内呈明显的时间效应关系，而GSH含量在6h后明显降低。结论 100mmol／L乙醇急性暴露可导致人原代培养肝细胞明显的氧化损伤，这种损伤与CYP 2E1活性的变化直接相关。

The relationship between CYP 2E1 activity and oxidative damage after acute ethanol exposure in human primary hepatocytes

Objective To study the oxidative damage to human primary cultured hepatocytes by acute ethanol exposure. Methods Primary hepatocytes were isolated and cultured from human liver tissue. After acute exposure to ethanol of different concentrations and different durations,CYP 2E1 activity in the hepatocytes was measured,meanwhile,glutathione (GSH),malondialdehyde (MDA) in the hepatocytes and aspartate transaminase (AST) in the supernatant were determined to indicate oxidative damage to the cells. Results There was a remarkable dose-and time-dependent relationship between ethanol exposure and CYP 2E1 activity in the human primary cultured hepatocytes. Moreover,there was a time-dependent increase in MDA content in hepatocytes and AST release in supernatants,while the GSH rose at the beginning and later declined significantly. Conclusion There was a markedly oxidative damage to human primary hepatocytes after acute ethanol exposure,and it is clearly related to the activity of CYP 2E1.