CYP2E1-dependent hepatotoxicity and oxidative damage after ethanol administration in human primary hepatocytes.
OBJECTIVE
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.


CONCLUSIONS
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.
Liu
Lie-Gang
LG
Department of Nutrition and Food Hygiene, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China. lgliu@mails.tjmu.edu.cn
Yan
Hong
H
Yao
Ping
P
Zhang
Wen
W
Zou
Li-Jun
LJ
Song
Fang-Fang
FF
Li
Ke
K
Sun
Xiu-Fa
XF
D002478:Cells, Cultured; D002492:Central Nervous System Depressants / Q000633:toxicity*; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D000431:Ethanol / Q000633:toxicity*; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology*; D006801:Humans; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology
D002492:Central Nervous System Depressants; D000431:Ethanol; D019392:Cytochrome P-450 CYP2E1