Document Detail


Inflammatory levels of nitric oxide inhibit airway epithelial cell migration by inhibition of the kinase ERK1/2 and activation of hypoxia-inducible factor-1 alpha.
MedLine Citation:
PMID:  18424783     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Increased synthesis of NO during airway inflammation, caused by induction of nitric-oxide synthase 2 in several lung cell types, may contribute to epithelial injury and permeability. To investigate the consequence of elevated NO production on epithelial function, we exposed cultured monolayers of human bronchial epithelial cells to the NO donor diethylenetriaamine NONOate. At concentrations generating high nanomolar levels of NO, representative of inflammatory conditions, diethylenetriaamine NONOate markedly reduced wound closure in an in vitro scratch injury model, primarily by inhibiting epithelial cell migration. Analysis of signaling pathways and gene expression profiles indicated a rapid induction of the mitogen-activated protein kinase phosphatase (MPK)-1 and decrease in extracellular signal-regulated kinase (ERK)1/2 activation, as well as marked stabilization of hypoxia-inducible factor (HIF)-1alpha and activation of hypoxia-responsive genes, under these conditions. Inhibition of ERK1/2 signaling using U0126 enhanced HIF-1alpha stabilization, implicating ERK1/2 dephosphorylation as a contributing mechanism in NO-mediated HIF-1alpha activation. Activation of HIF-1alpha by the hypoxia mimic cobalt chloride, or cell transfection with a degradation-resistant HIF-1alpha mutant construct inhibited epithelial wound repair, implicating HIF-1alpha in NO-mediated inhibition of cell migration. Conversely, NO-mediated inhibition of epithelial wound closure was largely prevented after small interfering RNA suppression of HIF-1alpha. Finally, NO-mediated inhibition of cell migration was associated with HIF-1alpha-dependent induction of PAI-1 and activation of p53, both negative regulators of epithelial cell migration. Collectively, our results demonstrate that inflammatory levels of NO inhibit epithelial cell migration, because of suppression of ERK1/2 signaling, and activation of HIF-1alpha and p53, with potential consequences for epithelial repair and remodeling during airway inflammation.
Authors:
Peter F Bove; Milena Hristova; Umadevi V Wesley; Nels Olson; Karen M Lounsbury; Albert van der Vliet
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Publication Detail:
Type:  Journal Article     Date:  2008-04-18
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  283     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2008 Jun 
Date Detail:
Created Date:  2008-06-23     Completed Date:  2008-08-18     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  17919-28     Citation Subset:  IM    
Affiliation:
Department of Pathology, College of Medicine, University of Vermont, Burlington, VT 05405, USA.
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MeSH Terms
Descriptor/Qualifier:
Cell Movement
Enzyme Activation
Epithelial Cells / cytology*
Gene Expression Regulation, Enzymologic*
Humans
Hypoxia-Inducible Factor 1, alpha Subunit / biosynthesis*
Inflammation*
Mitogen-Activated Protein Kinase 1 / metabolism*
Mitogen-Activated Protein Kinase 3 / metabolism*
Models, Biological
Nitric Oxide / metabolism*
RNA, Small Interfering / metabolism
Tumor Suppressor Protein p53 / metabolism*
Vascular Endothelial Growth Factor A / metabolism
Wound Healing
Chemical
Reg. No./Substance:
0/HIF1A protein, human; 0/Hypoxia-Inducible Factor 1, alpha Subunit; 0/RNA, Small Interfering; 0/Tumor Suppressor Protein p53; 0/Vascular Endothelial Growth Factor A; 10102-43-9/Nitric Oxide; EC 2.7.11.24/Mitogen-Activated Protein Kinase 1; EC 2.7.11.24/Mitogen-Activated Protein Kinase 3
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