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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 2 5～ 10 0mmol L乙醇作用于人原代肝细胞 9h及 10 0mmol L乙醇作用于人原代肝细胞 0～ 2 4h后 ,检测人原代肝细胞中CYP 2E1的含量 ,并研究 10 0mmol L乙醇作用于人原代肝细胞 0～ 2 4h后 ,天冬氨酸转胺酶 (aspartatetransaminase ,AST)的释放量及肝细胞中谷胱甘肽 (Glutathione ,GSH)、丙二醛 (Malondialdehyde ,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 10 0mmol L乙醇作用下 ,AST和MDA明显升高 ,在 0～ 2 4h内呈明显的时间效应关系 ,而GSH含量在 6h后明显降低。结论 10 0mmol 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。
Objective To investigate the toxic effects and oxidative damage of CYP 2E1 in human primary cultured hepatocytes under acute alcohol exposure. Methods Human primary hepatocytes were isolated and cultured. Human primary hepatocytes were treated with 20-50 mmol L ethanol for 9 h and 10 mmol L ethanol for 0-24 h. CYP 2E1 and the release of aspartate transaminase (AST) and the content of glutathione (GSH) in liver cells after 0 ~ 24 h of human primary hepatocytes were treated with 100 mmol L ethanol ), Malondialdehyde (MDA) content. Results Acute alcohol exposure led to the increase of CYP 2E1 release in human primary hepatocytes, and showed a dose-response and time-dependent effect. AST and MDA were significantly increased in the presence of 10 mmol L ethanol, ranging from 0 to 24 h Obvious time effect, while GSH content decreased significantly after 6h. Conclusion Acute 10 mmol L ethanol exposure can lead to obvious oxidative damage of human primary culture hepatocytes, which is directly related to the change of CYP 2E1 activity.