title: Cytochrome P4502E1 inhibitor, chlormethiazole, decreases lipopolysaccharide-induced inflammation in rat Kupffer cells with ethanol treatment.
abstract: OBJECTIVE
  To investigate the role of Cytochrome P4502E1 in sensitizing Kupffer cells to lipopolysaccharide (LPS)-mediated inflammation after ethanol induction.
  
  
  METHODS
  Sprague-Dawley 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-κB) p65 protein and tumor necrosis factor (TNF)-α mRNA were measured. In vitro, isolated Kupffer 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 Kupffer cell to LPS. The effect of LPS alone was tested in controlled Kupffer cells without ethanol. TNF-α, nuclear NF-κB p65 and cytoplasm IκB-α were monitored for all groups.
  
  
  RESULTS
  Ethanol feeding increased hepatic CYP2E1 level, nuclear accumulation of NF-κB p65 and TNF-α expression in rats. These changes were inhibited by CMZ supplementation. In cultured Kupffer 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-κB p65 and less cytoplasm IκB-α under LPS stimuli. CMZ abolished the effects of ethanol on LPS-stimulated NF-κB translocation and TNF-α generation in Kupffer cells.
  
  
  CONCLUSIONS
  In cultured Kupffer cell, using CMZ as inhibitor, ethanol-induced CYP2E1 overexpression was proved to contribute to the sensitization of Kupffer cells to LPS stimuli, with amplification of ROS production and activation of NF-κB, resulting in increased TNF-α production.
authors: Guangdong Provincial Key Laboratory of Food, Department of Nutrition, School of Public Health, Sun Yat-Sen University (Northern Campus), Guangzhou, China; Nutrition and Cancer Biology Laboratory, Jean Mayer USDA Human Nutrition Research Center on Aging, Tufts University, Boston, Massachusetts, USA.
authors: Qinyuan
authors: Q
authors: Ye
authors: Xiangdong
authors: X
authors: Wang
authors: Qing
authors: Q
authors: Wang
authors: Min
authors: M
authors: Xia
authors: Yanna
authors: Y
authors: Zhu
authors: Fuzhi
authors: F
authors: Lian
authors: Wenhua
authors: W
authors: Ling
keywords: Cytochrome P4502E1; Kupffer cell; alcoholic liver disease; chlormethiazole; lipopolysaccharide; nuclear factor-κB