Nitric oxide inhibits enterocyte migration through activation of RhoA-ATPase in a SHP-2-dependent manner

作者：Cetin Selma; Leaphart Cynthia L; Li Jun; Ischenko Irene; Hayman Michael; Upperman Jeffrey; Zamora Ruben; Watkins Simon; Ford Henri R; Wang James; Hackam David J^*

来源：American Journal of Physiology - Gastrointestinal and Liver Physiology, 2007, 292(5): G1347-G1358.

DOI：10.1152/ajpgi.00375.2006

• intestinal inflammation
• restitution
• epithelial barrier
• necrotizing enterocolitis
• bacterial translocation

Diseases of intestinal inflammation like necrotizing enterocolitis ( NEC) are associated with impaired epithelial barrier integrity and the sustained release of intestinal nitric oxide ( NO). NO modifies the cytoskeletal regulator RhoA-GTPase, suggesting that NO could affect barrier healing by inhibiting intestinal restitution. We now hypothesize that NO inhibits enterocyte migration through RhoA- GTPase and sought to determine the pathways involved. The induction of NEC was associated with increased enterocyte NO release and impaired migration of bromodeoxyuridine-labeled enterocytes from terminal ileal crypts to villus tips. In IEC- 6 enterocytes, NO significantly inhibited enterocyte migration and activated RhoA- GTPase while increasing the formation of stress fibers. In parallel, exposure of IEC- 6 cells to NO increased the phosphorylation of focal adhesion kinase ( pFAK) and caused a striking increase in cell- matrix adhesiveness, suggesting a mechanism by which NO could impair enterocyte migration. NEC was associated with increased expression of pFAK in the terminal ileal mucosa of wild- type mice and a corresponding increase in disease severity compared with inducible NO synthase knockout mice, confirming the dependence of NO for FAK phosphorylation in vivo and its role in the pathogenesis of NEC. Strikingly, inhibition of the protein tyrosine phosphatase SHP- 2 in IEC- 6 cells prevented the activation of RhoA by NO, restored focal adhesions, and reversed the inhibitory effects of NO on enterocyte migration. These data indicate that NO impairs mucosal healing by inhibiting enterocyte migration through activation of RhoA in a SHP- 2- dependent manner and support a possible role for SHP- 2 as a therapeutic target in diseases of intestinal inflammation like NEC.

• 出版日期2007-5