Nitric oxide induces HIF-1α stabilization and expression of intestinal trefoil factor in the damaged rat jejunum and modulates ulcer healing

J Gastroenterol. 2011 May;46(5):565-76. doi: 10.1007/s00535-011-0374-1. Epub 2011 Feb 9.


Background: The induction of intestinal trefoil factor (ITF) has been reported to depend on hypoxia-inducible factor-1 (HIF-1). Nitric oxide modulates HIF-1 activity. The present study aims to analyze the role of nitric oxide in jejunum damage induced by indomethacin and its ability to modulate epithelial function through the expression of ITF.

Methods: Rats received indomethacin (7.5 mg/kg, s.c., twice), and a time course analysis of damage was performed (24-96 h after the first administration). In these animals, the role of nitric oxide was analyzed by using 1400W, a selective iNOS activity inhibitor (5 mg/kg, i.p./day), on: (1) intestinal damage, (2) ulcer healing, (3) the presence of nitrated proteins in the jejunum and (4) the protein expression of inducible nitric oxide synthase (iNOS), HIF-1α and ITF.

Results: Indomethacin induced damage in the jejunum that was apparent at 24 h and peaked at 48-72 h. An increase in iNOS, HIF-1α, ITF and nitrated proteins was observed in the injured jejunum. Immunoprecipitation of HIF-1α allowed determination of the nitration/nitrosylation of this protein by using nitrotyrosine and nitrocysteine antibodies. Blockade of iNOS activity did not significantly modify damage or iNOS expression, but did significantly impede ITF induction, HIF-1α stabilization and HIF-1α detection with antibodies against nitrated proteins. In parallel to these results, pre-treatment with 1400W delayed the healing of the ulcer provoked by indomethacin.

Conclusions: These results suggest that iNOS-derived NO is involved in HIF-1α stabilization, probably through S-nitrosylation, and ITF expression in goblet cells of the damaged jejunum of indomethacin-treated rats and mediates ulcer healing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Inflammatory Agents, Non-Steroidal / toxicity
  • Goblet Cells / metabolism
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism*
  • Imines / pharmacology
  • Immunoprecipitation
  • Indomethacin / toxicity*
  • Intestinal Mucosa / metabolism
  • Jejunum / drug effects
  • Jejunum / pathology
  • Male
  • Nitric Oxide / metabolism*
  • Nitric Oxide Synthase Type II / metabolism
  • Peptic Ulcer / chemically induced
  • Peptic Ulcer / pathology
  • Peptides / metabolism*
  • Rats
  • Rats, Sprague-Dawley
  • Time Factors
  • Trefoil Factor-2


  • Anti-Inflammatory Agents, Non-Steroidal
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Imines
  • N-((3-(aminomethyl)phenyl)methyl)ethanimidamide
  • Peptides
  • Trefoil Factor-2
  • Nitric Oxide
  • Nitric Oxide Synthase Type II
  • Indomethacin