Inactivation of HNF4a mediates alcohol-induced downregulation of intestinal tight junction proteins.pdf

Inactivation of hepatocyte nuclear factor-4
mediates alcohol-induced downregulation of intestinal tight junction proteins Wei Zhong,1,5 Yantao Zhao,1,6 Craig J. McClain,1,2,3,4 Y. James Kang,2,3 and Zhanxiang Zhou1,3 Departments of 1Medicine and 2Pharmacology Toxicology, University of Louisville School of Medicine and 3University of Louisville Alcohol Research Center, 4Louisville Veterans’ Affairs Medical Center, Louisville, Kentucky; 5College of Veterinary Medicine, China Agricultural University, Beijing; and 6College of Animal Sciences and Veterinary Medicine, Agricultural University of Hebei, Baoding, China Submitted 22 December 2009; accepted in final form 18 June 2010 Zhong W, Zhao Y, McClain CJ, Kang YJ, Zhou Z. Inactivation of hepatocyte nuclear factor-4
mediates alcohol-induced downregu- lation of intestinal tight junction proteins. Am J Physiol Gastrointest Liver Physiol 299: G643–G651, 2010. First published June 24, 2010; doi:10.1152/ajpgi.00515.2009.—Chronic alcohol exposure has been shown to increase the gut permeability in the distal intestine, in part, through induction of zinc deficiency. The present study evaluated the molecular mechanisms whereby zinc deficiency mediates alcohol- induced intestinal barrier dysfunction. Examination of zinc finger transcription factors in the gastrointestinal tract of mice revealed a prominent distribution of hepatocyte nuclear factor-4
(HNF-4
). HNF-4
exclusively localizes in the epithelial nuclei and exhibited an increased abundance in mRNA and protein levels in the distal intes- tine. Chronic alcohol exposure to mice repressed the HNF-4
gene expression in the ileum and reduced the protein level and DNA binding activity of HNF-4
in all of the intestinal segments with the most remarkable changes in the ileum. Chronic alcohol exposure also decreased the mRNA levels of tight junction proteins, particularly in the ileum. Caco-2 cell culture studies were conducted to determine the role of HNF-4
in regulation of t