TNF-alpha modulates iNOS expression in an experimental rat model of indomethacin-induced jejunoileitis

Mol Cell Biochem. 2010 Mar;336(1-2):17-24. doi: 10.1007/s11010-009-0259-2. Epub 2009 Oct 3.


Multiple mucosal immune factors, such as TNF-alpha and IL-1beta, are thought to be key mediators involved in inflammatory bowel disease. We evaluated the role of the pro-inflammatory cytokine TNF-alpha on nitric oxide synthase (NOS) expression in indomethacin-induced jejunoileitis in rats. Jejunoileitis was induced in rats with subcutaneous injections of indomethacin (7.5 mg/kg) 24 h apart for two consecutive days, and animals were randomized into four groups. Group 1 received only indomethacin. Group 2 was treated with a daily dose of phosphodiesterase (PDE) inhibitor (theophylline or pentoxifylline) by oral gavage for 2 days before and 4 days after indomethacin. Group 3 received a single dose of anti-TNF-alpha monoclonal antibody (TNF-Ab, IP) 30 min before indomethacin. Group 4 was treated with 1 h hyperbaric oxygenation (HBO(2)) for 5 days after indomethacin. Rats were sacrificed at 12 h or 4 days after final indomethacin injection. PDE inhibitor, TNF-Ab, or HBO(2) treatment significantly decreased indomethacin-induced ulceration, myeloperoxidase activity, and disease activity index. Although indomethacin significantly increased serum TNF-alpha and nitrate/nitrite (NOx) concentrations above control values at 12 h, inducible NOS (iNOS) expression was detected only at day 4. Serum IL-1beta levels did not change at 12 h but increased 4-fold after 4 days. Indomethacin had no effect on constitutive NOS. Treatment with PDE inhibitor, TNF-Ab, or HBO(2) significantly reduced serum/tissue TNF-alpha, IL-1beta, NOx, and iNOS expression. Our data show TNF-alpha plays an early pro-inflammatory role in indomethacin-induced jejunoileitis. Additionally, down-regulation of NOx by PDE inhibitors, TNF-Ab, or HBO(2) suggests that TNF-alpha modulates iNOS expression.

MeSH terms

  • Animals
  • Dose-Response Relationship, Drug
  • Enteritis / blood
  • Enteritis / chemically induced
  • Enteritis / metabolism*
  • Hyperbaric Oxygenation
  • Ileitis / blood
  • Ileitis / chemically induced
  • Ileitis / metabolism*
  • Ileum / enzymology
  • Ileum / metabolism
  • Indomethacin / toxicity*
  • Interleukin-1beta / blood
  • Interleukin-1beta / metabolism
  • Intestinal Mucosa / enzymology
  • Intestinal Mucosa / metabolism
  • Jejunal Diseases / blood
  • Jejunal Diseases / chemically induced
  • Jejunal Diseases / metabolism*
  • Jejunum / enzymology
  • Jejunum / metabolism
  • Male
  • Nitrates / blood
  • Nitric Oxide Synthase Type II / metabolism*
  • Nitrites / blood
  • Peroxidase / metabolism
  • Phosphodiesterase Inhibitors / pharmacology
  • Random Allocation
  • Rats
  • Rats, Sprague-Dawley
  • Severity of Illness Index
  • Specific Pathogen-Free Organisms
  • Time Factors
  • Tumor Necrosis Factor-alpha / antagonists & inhibitors
  • Tumor Necrosis Factor-alpha / blood
  • Tumor Necrosis Factor-alpha / physiology*


  • Interleukin-1beta
  • Nitrates
  • Nitrites
  • Phosphodiesterase Inhibitors
  • Tumor Necrosis Factor-alpha
  • Peroxidase
  • Nitric Oxide Synthase Type II
  • Indomethacin