Role for intestinal CYP2E1 in alcohol-induced circadian gene-mediated intestinal hyperpermeability

Christopher B. Forsyth, Robin M. Voigt, Maliha Shaikh, Yueming Tang, Arthur I. Cederbaum, Fred W. Turek, Ali Keshavarzian

Research output: Contribution to journalArticlepeer-review

54 Scopus citations


We have shown that alcohol increases Caco-2 intestinal epithelial cell monolayer permeability in vitro by inducing the expression of redoxsensitive circadian clock proteins CLOCK and PER2 and that these proteins are necessary for alcohol-induced hyperpermeability. We hypothesized that alcohol metabolism by intestinal Cytochrome P450 isoform 2E1 (CYP2E1) could alter circadian gene expression (Clock and Per2), resulting in alcohol-induced hyperpermeability. In vitro Caco-2 intestinal epithelial cells were exposed to alcohol, and CYP2E1 protein, activity, and mRNA were measured. CYP2E1 expression was knocked down via siRNA and alcohol-induced hyperpermeability, and CLOCK and PER2 protein expression were measured. Caco-2 cells were also treated with alcohol or H2O2 with or without N-acetylcysteine (NAC) anti-oxidant, and CLOCK and PER2 proteins were measured at 4 or 2 h. In vivo Cyp2e1 protein and mRNA were also measured in colon tissue from alcohol-fed mice. Alcohol increased CYP2E1 protein by 93% and enzyme activity by 69% in intestinal cells in vitro. Alcohol feeding also increased mouse colonic Cyp2e1 protein by 73%. mRNA levels of Cyp2e1 were not changed by alcohol in vitro or in mouse intestine. siRNA knockdown of CYP2E1 in Caco-2 cells prevented alcohol-induced hyperpermeability and induction of CLOCK and PER2 proteins. Alcohol-induced and H2O2-induced increases in intestinal cell CLOCK and PER2 were significantly inhibited by treatment with NAC. We concluded that our data support a novel role for intestinal CYP2E1 in alcohol-induced intestinal hyperpermeability via a mechanism involving CYP2E1- dependent induction of oxidative stress and upregulation of circadian clock proteins CLOCK and PER2.

Original languageEnglish
Pages (from-to)G185-G195
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Issue number2
StatePublished - 15 Jul 2013


  • Clock genes
  • Cytochrome P450 isoform 2E1
  • Ethanol
  • Leaky gut
  • Oxidative stress


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