Increased expression and cellular localization of inducible nitric oxide synthase and cyclooxygenase 2 in Helicobacter pylori gastritis

doi:10.1016/S0016-5085(99)70496-8

Gastroenterology

Volume 116, Issue 6, June 1999, Pages 1319-1329

Presented in part at Digestive Diseases Week in May 1995 (San Diego, CA) and in May 1997 (Washington, D.C.).

https://doi.org/10.1016/S0016-5085(99)70496-8 Get rights and content

Abstract

Background & Aims: Inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 are important regulators of mucosal inflammation and epithelial cell growth. To determine the role of iNOS and COX-2 in Helicobacter pylori–induced tissue injury, we compared their gene expression in H. pylori–induced gastritis with that in normal gastric mucosa and in non–H. pylori gastritis. Methods: In 43 patients, we assessed H. pylori infection status, histopathology, messenger RNA (mRNA) and protein expression, and cellular localization of iNOS and COX-2. Results: By reverse-transcription polymerase chain reaction (RT-PCR), antral iNOS and COX-2 mRNA expression was absent to low in normal mucosa (n = 10), significantly increased in H. pylori–negative gastritis (n = 13), and even more markedly increased in H. pylori–positive gastritis (n = 20). Increased iNOS and COX-2 levels were confirmed by Northern and Western blot analysis and were both greater in the gastric antrum than in the gastric body of infected patients. Immunohistochemistry also showed increased expression of both genes in H. pylori gastritis: iNOS protein was detected in epithelium, endothelium, and lamina propria inflammatory cells, and COX-2 protein localized to mononuclear and fibroblast cells in the lamina propria. Conclusions: iNOS and COX-2 are induced in H. pylori–positive gastritis and thus may modulate the inflammation and alterations in epithelial cell growth that occur in this disease. Higher levels of iNOS and COX-2 in H. pylori–positive vs. –negative gastritis and in gastric antrum, where bacterial density is greatest, suggest that expression of these genes is a direct response to H. pylori infection.

GASTROENTEROLOGY 1999;116:1319-1329

Section snippets

Tissue samples

All tissues in this study were obtained from endoscopic biopsy specimens from patients attending either the Baltimore Veterans Affairs or University of Maryland Medical Center. Patients presenting for esophagogastroduodenoscopy for clinical indications were considered for inclusion in the study. Patients previously treated for H. pylori infection were excluded, as were patients admitting to active use of nonsteroidal anti-inflammatory drugs (NSAIDs) at least once per week. All patients signed

Clinical and histological characteristics

Tissues from 43 patients were analyzed. Twenty were classified as H. pylori–positive. All had the presence of H. pylori detected on histology; gastritis scores are shown in Table 1. Of these patients, 19 of 20 had positive CLOtest results, and the one patient with negative CLOtest results had a positive serology. Of the remaining 23 patients with no histological evidence of H. pylori and negative results of CLOtest and H. pylori serology, 10 had normal gastric histology and 13 had evidence of

Discussion

The above findings suggest that iNOS and COX-2 are important factors in the pathogenesis of H. pylori–induced gastritis. Expression of both genes was markedly up-regulated at the mRNA and protein levels in gastric mucosa from patients with H. pylori–positive gastritis compared with normal mucosa. Overall, expression levels were significantly higher in tissues from H. pylori–positive gastritis than in H. pylori–negative gastritis, suggesting that induction of iNOS and COX-2 may be a specific

Acknowledgements

Drs. Fu and Ramanujam contributed equally to this work.

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^☆: Address requests for reprints to: Keith T. Wilson, M.D., Division of Gastroenterology, University of Maryland Medical Systems, Room N3W62, 22 South Greene Street, Baltimore, Maryland 21201. e-mail: [email protected]; fax: (410) 328-8315.

^☆☆: Supported by National Institutes of Health grants K08-DK02469 (to K.T.W.), R01-CA67497 (to S.J.M. and K.T.W.), and R01-DK53620 (to S.P.J., S.J.M., and K.T.W.); by the Robert and Sally D. Funderburg Award in Gastric Cancer Biology (to S.J.M.); by University of Maryland School of Medicine (K.T.W.); the Crohn's and Colitis Foundation (K.T.W.); Astra Merck (G.T.F. and K.T.W.); and the Office of Medical Research, Department of Veterans Affairs (S.J.M.).

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Alimentary TractIncreased expression and cellular localization of inducible nitric oxide synthase and cyclooxygenase 2 in Helicobacter pylori gastritis☆,☆☆

Abstract

Section snippets

Tissue samples

Clinical and histological characteristics

Discussion

Acknowledgements

Biochem Biophys Res Commun

Gastroenterology

Gastroenterology

Gastroenterology

Gastroenterology

Lancet

Gastroenterology

J Biol Chem

Cell

Gastroenterology

Cell

Gastroenterology

Gastroenterology

Gastroenterology

Gastrointest Endosc

Anal Biochem

J Biol Chem

J Biol Chem

J Biol Chem

Gastroenterology

Gastroenterology

Lancet

Gastroenterology

Gastroenterology

Gastroenterology

Cloning and characterization of inducible nitric oxide synthase from mouse macrophages

Science

Cloned and expressed macrophage nitric oxide synthase contrasts with the brain enzyme

Proc Natl Acad Sci USA

Cloning and characterization of a growth factor–inducible cyclo-oxygenase gene from rat intestinal epithelial cells

Am J Physiol

Nitric oxide synthase activity in ulcerative colitis and Crohn's disease

Lancet

Amelioration of chronic ileitis by nitric oxide synthase inhibition

J Pharmacol Exp Ther

Co-expression of inducible nitric oxide synthase (iNOS), cyclo-oxygenase (COX-2), and TGF-β in rat models of colitis (abstr)

Gastroenterology

Exacerbation of inflammation-associated colonic injury in rat through inhibition of cyclo-oxygenase-2

J Clin Invest

Increased expression of inducible nitric oxide synthase and cyclo-oxygenase-2 in Barrett's esophagus and associated adenocarcinomas

Cancer Res

Alimentary Tract
Increased expression and cellular localization of inducible nitric oxide synthase and cyclooxygenase 2 in Helicobacter pylori gastritis☆,☆☆