Excessive secretion of interleukin-8 (IL-8), a pro-inflammatory chemokine secreted by activated endothelium during acute inflammation, causes neutrophil-induced vascular injury during processes such as bacterial sepsis, multiple trauma, or reperfusion of ischemic tissue. Nitric oxide (NO), a key signaling molecule, exhibits anti-inflammatory properties where endothelial activation and extreme IL-8 secretion are present. Research performed by the sponsor shows that NO may exert different regulatory effects on IL-8 expression according to the state of endothelial cell activation. This application's broad objective is to determine NO's role in regulating vascular endothelial responses to acute inflammation examining three specific aims: 1) to characterize the regulation of IL-8 gene expression by the transcription factors AP-1 and NF-kappaB; 2) to determine the effect of NO on the signaling pathway involved in the activation of AP-1 by inflammatory stimuli such as tumor necrosis factor-alpha (TNF-alpha); 3) to determine the effect of NO on the regulatory proteins involved in the activation of NF- kappaB by inflammatory stimuli such as TNF-alpha and interleukin- 1. Molecular techniques such as polymerase chain reaction, transient transfection, dual luciferase reporter assays, site directed mutagenesis, gel mobility shift assays and kinase assays will be used to accomplish these aims. Existing knowledge concerning NO's role in regulating inflammation is incomplete, yet NO is currently under study in human clinical trials as an adjunctive therapy for acute inflammatory vascular injury of the lung. This research will provide information essential to more completely understand the role of NO in the inflammatory process.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32HL010355-02
Application #
6388774
Study Section
Lung Biology and Pathology Study Section (LBPA)
Program Officer
Schucker, Beth
Project Start
2001-07-01
Project End
Budget Start
2001-07-01
Budget End
2002-06-30
Support Year
2
Fiscal Year
2001
Total Cost
$41,996
Indirect Cost
Name
Virginia Commonwealth University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
City
Richmond
State
VA
Country
United States
Zip Code
23298
Natarajan, Ramesh; Jones, Drew G; Fisher, Bernard J et al. (2005) Hypoxia inducible factor-1: regulation by nitric oxide in posthypoxic microvascular endothelium. Biochem Cell Biol 83:597-607
Natarajan, Ramesh; Fisher, Bernard J; Fowler 3rd, Alpha A (2003) Regulation of hypoxia inducible factor-1 by nitric oxide in contrast to hypoxia in microvascular endothelium. FEBS Lett 549:99-104
Natarajan, Ramesh; Fisher, Bernard J; Jones, Drew G et al. (2002) Reoxygenating microvascular endothelium exhibits temporal dissociation of NF-kappaB and AP-1 activation. Free Radic Biol Med 32:1033-45
Natarajan, Ramesh; Fisher, Bernard J; Jones, Drew G et al. (2002) Atypical mechanism of NF-kappaB activation during reoxygenation stress in microvascular endothelium: a role for tyrosine kinases. Free Radic Biol Med 33:962
Benjamin, I J; Shelton, J; Garry, D J et al. (1997) Temporospatial expression of the small HSP/alpha B-crystallin in cardiac and skeletal muscle during mouse development. Dev Dyn 208:75-84