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Cytochrome P4502E1 inhibitor, chlormethiazole, decreases lipopolysaccharide‐induced inflammation in rat K upffer cells with ethanol treatment
Author(s) -
Ye Qinyuan,
Wang Xiangdong,
Wang Qing,
Xia Min,
Zhu Yanna,
Lian Fuzhi,
Ling Wenhua
Publication year - 2013
Publication title -
hepatology research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.123
H-Index - 75
eISSN - 1872-034X
pISSN - 1386-6346
DOI - 10.1111/hepr.12063
Subject(s) - cyp2e1 , lipopolysaccharide , tumor necrosis factor alpha , kupffer cell , chemistry , ethanol , nf κb , sensitization , inflammation , microbiology and biotechnology , in vitro , biochemistry , endocrinology , biology , apoptosis , immunology , microsome
Aim To investigate the role of C ytochrome P4502E1 in sensitizing K upffer cells to lipopolysaccharide ( LPS )‐mediated inflammation after ethanol induction. Methods S prague– D awley rats were fed a liquid ethanol diet, control diet or ethanol diet supplemented with CYP2E1 inhibitor, chlormethiazole ( CMZ ), for 4 weeks. Hepatic CYP2E1 protein, nuclear factor‐kappa B ( NF ‐κ B ) p65 protein and tumor necrosis factor ( TNF )‐α m RNA were measured. In vitro , isolated K upffer cells from control rats were exposed to ethanol with different CMZ concentration; CYP2E1 expression and reactive oxygen species ( ROS ) generation were compared. The identified CMZ concentration was further utilized to evaluate the role of CYP2E1 on the sensitization of ethanol‐induced K upffer cell to LPS . The effect of LPS alone was tested in controlled K upffer cells without ethanol. TNF ‐α, nuclear NF ‐κ B p65 and cytoplasm I κ B ‐α were monitored for all groups. Results Ethanol feeding increased hepatic CYP2E1 level, nuclear accumulation of NF ‐κ B p65 and TNF ‐α expression in rats. These changes were inhibited by CMZ supplementation. In cultured K upffer cells, increased CYP2E1 content and ROS production by in vitro ethanol induction were dose‐dependently inhibited by CMZ . Compared with LPS alone, the ethanol induction group produced significantly more TNF ‐α, nuclear NF ‐κ B p65 and less cytoplasm I κ B ‐α under LPS stimuli. CMZ abolished the effects of ethanol on LPS ‐stimulated NF ‐κ B translocation and TNF ‐α generation in K upffer cells. Conclusion In cultured K upffer cell, using CMZ as inhibitor, ethanol‐induced CYP2E1 overexpression was proved to contribute to the sensitization of K upffer cells to LPS stimuli, with amplification of ROS production and activation of NF ‐κ B , resulting in increased TNF ‐α production.