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Am J Physiol Cell Physiol (October 10, 2001). doi:10.1152/ajpcell.00295.2001
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Articles in PresS, published online ahead of print October 10, 2001
Am J Physiol Cell Physiol, 10.1152/ajpcell.00295.2001
Submitted on June 28, 2001
Accepted on October 5, 2001

Peroxynitrite Increases iNOS Through NF{kappa}B and Decreases Prostacyclin Synthase in Endothelial Cells

Christy-Lynn M Cooke1 and Sandra T Davidge1*

1 Perinatal Research Centre, University of Alberta, Edmonton, Alberta, Canada

* To whom correspondence should be addressed. E-mail: sandra.davidge{at}ualberta.ca.

Peroxynitrite, a marker of oxidative stress, is elevated in conditions associated with vascular endothelial cell dysfunction, such as atherosclerosis, preeclampsia and diabetes. However, the effects of peroxynitrite on endothelial cell function are not clear. Endothelial-derived enzymes NOS and PGHS mediate vascular reactivity and contain oxidant-sensitive isoforms (iNOS and PGHS-2) which can be induced by NF{kappa}B activation. We investigated the effect(s) of peroxynitrite on NOS and PGHS pathways in endothelial cells. We hypothesized that peroxynitrite will increase levels of iNOS and PGHS-2, through activation of NF{kappa}B. Western immunoblots of endothelial cells show that SIN-1 (0.5 mM), a peroxynitrite donor, increased iNOS protein mass, which can be inhibited by PDTC (NF{kappa}B inhibitor) (167±24.2% vs. 78±19%, P<0.05, n=6). SIN-1 treatment also significantly increased NF{kappa}B translocation into endothelial cell nuclei (135±10%, P<0.05). eNOS, PGHS-1 and PGHS-2 protein levels were not altered by SIN-1. However, prostacyclin synthase protein mass, but not mRNA, was significantly reduced in SIN-1 treated endothelial cells (78±8.9%, P<0.05). Our results illustrate novel mechanisms through which peroxynitrite may modulate vascular endothelial function.




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