Bacterial induction of inducible nitric oxide synthase in cultured human intestinal epithelial cells

Gastroenterology
A L SalzmanC Szabó

Abstract

Enterocytes play a major role in the mucosa as a source of proinflammatory cytokines and cytotoxins. We tested the hypothesis that bacteria induce expression of the inducible nitric oxide synthase (iNOS) in cultured human enterocytes. DLD-1 and Caco-2BBe cell monolayers exposed to Salmonella dublin were analyzed for iNOS up-regulation and nitric oxide production (NOx) in the presence of various proinflammatory cytokines. S. dublin augmented NOx in interferon gamma (IFN-gamma)-primed cells but had no independent effect on iNOS expression. S. dublin-induced NOx was not mediated by endotoxin and was augmented by an enteroinvasive phenotype. In DLD-1 cells, S. dublin-mediated NOx was blocked by inhibitors of nuclear factor kappa B (NF-kappa B) and tyrosine kinase activation and was steroid resistant. Cis-acting elements in the human iNOS promoter responsive to endotoxin and S. dublin stimulation of IFN-gamma-treated DLD-1 cells were identified between 10.9 and 8.7 kilobases upstream of the transcription initiation site. S. dublin alters the regulation of iNOS messenger RNA in IFN-gamma-treated intestinal epithelial cells via a steroid-resistant pathway involving NF-kappa B and tyrosine kinase activity. Because bacterial interaction...Continue Reading

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