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B and decreases
prostacyclin synthase in endothelial cells
Perinatal Research Centre, Departments of Obstetrics/Gynecology and Physiology, University of Alberta, Edmonton, Alberta, Canada T6G 2S2
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. The endothelium-derived enzymes nitric oxide synthase (NOS)
and prostaglandin H synthase (PGHS) mediate vascular reactivity and
contain oxidant-sensitive isoforms (iNOS and PGHS-2) that can be
induced by nuclear factor (NF)-
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-B. Western immunoblots of
endothelial cells show that 3-morpholinosydnonimine (SIN-1; 0.5 mM), a
peroxynitrite donor, increased iNOS protein mass, which can be
inhibited by pyrroline dithiocarbamate (an NF-B inhibitor) (167 ± 24.2 vs. 78 ± 19%, P < 0.05, n = 6). SIN-1 treatment also significantly increased
NF-B translocation into endothelial cell nuclei (135 ± 10%,
P < 0.05). Endothelial NOS, 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.
oxidative stress; vascular function
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