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Am J Physiol Cell Physiol 284: C1397-C1404, 2003. First published February 12, 2003; doi:10.1152/ajpcell.00328.2002
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Vol. 284, Issue 6, C1397-C1404, June 2003

Neurotensin stimulates IL-8 expression in human colonic epithelial cells through Rho GTPase-mediated NF-kappa B pathways

Dezheng Zhao1, Sabina Kuhnt-Moore1, Huiyan Zeng2, Jack S. Wu1, Mary P. Moyer3, and Charalabos Pothoulakis1

1 Division of Gastroenterology, 2 Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02468; and 3 INCELL Corporation, San Antonio, Texas 78249

Neurotensin (NT), a neuropeptide highly expressed in the gastrointestinal tract, participates in the pathophysiology of intestinal inflammation. We recently showed that NT stimulates interleukin-8 (IL-8) expression in NCM460 nontransformed human colonic epithelial cells via both mitogen-activating protein kinase (MAPK)- and NF-kappa B-dependent pathways. However, the molecular mechanism by which NT induces expression of proinflammatory cytokines such as IL-8 has not been investigated. In this study we show that inhibition of endogenous Rho family proteins (RhoA, Rac1, and Cdc42) by their respective dominant negative mutants inhibits NT-induced IL-8 protein production and promoter activity. Western blot experiments demonstrated that NT strongly activated RhoA, Rac1, and Cdc42. Overexpression of the dominant negative mutants of RhoA, Rac1, and Cdc42 significantly inhibited NT-induced NF-kappa B-dependent reporter gene expression and NF-kappa B DNA binding activity. NT also stimulated p38 MAPK phosphorylation, and overexpression of dominant negative mutants of RhoA, Rac1, and Cdc42 did not significantly alter p38 and ERK1/2 phosphorylation in response to NT. Together, our findings indicate that NT-stimulated IL-8 expression is mediated via a Rho-dependent NF-kappa B-mediated pathway.

neuropeptide; inflammation; signal transduction; gene regulation


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