CYP2E1-dependent hepatotoxicity and oxidative damage after ethanol administration in human primary hepatocytes
CYP2E1-dependent hepatotoxicity and oxidative damage after ethanol administration in human primary hepatocytes
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DOI：
10.3748/wjg.v11.i29.4530
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发表时间：
2005-08-07
影响因子：
4.3
通讯作者：
Sun, Xiu-Fa
中科院分区：
文献类型：
--
作者：
Liu, Lie-Gang;Yan, Hong;Sun, Xiu-Fa
目的：目的：观察乙醇对人原代培养肝细胞氧化损伤与细胞色素P450 2 E1（CYP 2 E1）活性的关系，探讨抑制CYP 2 E1活性是否能减轻乙醇对肝细胞的损伤。分光光度计法检测细胞色素P450 2 E1活性，Western blot法检测细胞色素P450 2 E1蛋白表达。肝毒性通过测定肝细胞培养上清液中乳酸脱氢酶（LDH）和天冬氨酸转氨酶（AST）水平以及细胞内丙二醛（MDA）的形成来研究。100 mmol/L乙醇暴露后，CYP 2 E1蛋白表达呈时间依赖性增加。同时，乙醇暴露的肝细胞引起的细胞MDA水平，LDH，AST活性在上清液中的时间依赖性增加。结论：乙醇诱导的人原代肝细胞氧化损伤与CYP 2 E1活性呈正相关，抑制CYP 2 E1活性可部分减轻乙醇诱导的氧化损伤。(C)2005年WJG出版社和Elsevier Inc. All rights reserved.
AIM: To observe the relationship between ethanol-induced oxidative damage in human primary cultured hepatocytes and cytochrome P450 2E1 (CYP2E1) activity, in order to address if inhibition of CYP2E1 could attenuate ethanol-induced cellular damage.METHODS: The dose-dependent (25-100 mmol/L) and time-dependent (0-24 h) exposures of primary human cultured hepatocytes to ethanol were carried out. CYP2E1 activity and protein expression were detected by spectrophotometer and Western blot analysis respectively. Hepatotoxicity was investigated by determination of lactate dehydrogenase (LDH) and aspartate transaminase (AST) level in hepatocyte culture supernatants, as well as the intracellular formation of malondialdehyde (MDA).RESULTS: A dose-and time-dependent response between ethanol exposure and CYP2E1 activity in human hepatocytes was demonstrated. Moreover, there was a time-dependent increase of CYP2E1 protein after 100 mmol/L ethanol exposure. Meanwhile, ethanol exposure of hepatocytes caused a time-dependent increase of cellular MDA level, LDH, and AST activities in supernatants. Furthermore, the inhibitor of CYP2E1, diallyl sulfide (DAS) could partly attenuate the increases of MDA, LDH, and AST in human hepatocytes.CONCLUSION: A positive relationship between ethanol-induced oxidative damage in human primary cultured hepatocytes and CYP2E1 activity was exhibited, and the inhibition of CYP2E1 could partly attenuate ethanol-induced oxidative damage. (C) 2005 The WJG Press and Elsevier Inc. All rights reserved.