Rho kinase blockade prevents inflammation via nuclear factor kappa B inhibition: evidence in Crohn's disease and experimental colitis

Gastroenterology. 2003 May;124(5):1180-7. doi: 10.1016/s0016-5085(03)00283-x.

Abstract

Background & aims: Rho proteins are involved in the regulation of several cellular functions. Data from in vitro studies suggest that RhoA could be involved in the inflammatory response. We investigated the role of RhoA and its downstream effector Rho kinase in intestinal inflammation.

Methods: Activation of RhoA was assessed by pull-down assays. A specific inhibitor of Rho kinase, Y-27632, was used to examine the role of Rho kinase in inflammatory response in vivo and in vitro by molecular biology and by immunological and biochemical approaches.

Results: Increased activation of RhoA was found in inflamed intestinal mucosa of patients with Crohn's disease and of rats with 2,4,6-trinitrobenzene sulfonic acid-induced colitis. Oral administration of Y-27632 in rats significantly reduced the colonic inflammation. In vitro, activation of RhoA alone was sufficient to induce tumor necrosis factor production. Y-27632 inhibited production of tumor necrosis factor-alpha and interleukin-1 beta by lamina propria and peripheral blood mononuclear cells. Rho kinase inhibition prevented nuclear factor kappa B activation and I-kappa B phosphorylation and degradation. We showed that Rho kinase associates with and activates I-kappa B kinase alpha and that Y-27632 prevents I-kappa B kinase activation.

Conclusions: Our study provides the first evidence that Rho kinase activates I-kappa B kinase and, thus, nuclear factor kappa B, suggesting a key role of Rho kinase in inflammatory responses and intestinal inflammation. Specific inhibition of Rho kinase may be a promising approach for the treatment of patients with Crohn's disease.

Publication types

  • Clinical Trial
  • Controlled Clinical Trial
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Amides / administration & dosage*
  • Animals
  • Cells, Cultured
  • Colitis / drug therapy
  • Colitis / immunology
  • Colitis / metabolism
  • Crohn Disease / drug therapy*
  • Crohn Disease / immunology
  • Crohn Disease / metabolism
  • Cytokines / metabolism
  • Enzyme Inhibitors / administration & dosage*
  • Female
  • Humans
  • I-kappa B Kinase
  • Intestinal Mucosa / enzymology
  • Intestinal Mucosa / immunology
  • Intracellular Signaling Peptides and Proteins
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / immunology
  • Male
  • Middle Aged
  • NF-kappa B / antagonists & inhibitors*
  • Prospective Studies
  • Protein-Serine-Threonine Kinases / antagonists & inhibitors*
  • Protein-Serine-Threonine Kinases / metabolism
  • Pyridines / administration & dosage*
  • Rats
  • Rats, Sprague-Dawley
  • Trinitrobenzenesulfonic Acid
  • rho-Associated Kinases
  • rhoA GTP-Binding Protein / metabolism

Substances

  • Amides
  • Cytokines
  • Enzyme Inhibitors
  • Intracellular Signaling Peptides and Proteins
  • NF-kappa B
  • Pyridines
  • Y 27632
  • Trinitrobenzenesulfonic Acid
  • Protein-Serine-Threonine Kinases
  • rho-Associated Kinases
  • CHUK protein, human
  • I-kappa B Kinase
  • IKBKB protein, human
  • IKBKE protein, human
  • rhoA GTP-Binding Protein