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-induced impairment of
endothelium-dependent vasorelaxation in coronary arteries
Departments of Pharmacology and Toxicology and Physiology, Medical College of Wisconsin, Milwaukee, Wisconsin 53226
The present study tested the
hypothesis that ceramide, a sphingomylinase metabolite, serves as an
second messenger for tumor necrosis factor-
(TNF-) to stimulate
superoxide production, thereby decreasing endothelium-dependent
vasorelaxation in coronary arteries. In isolated bovine small coronary
arteries, TNF- (1 ng/ml) markedly attenuated vasodilator responses
to bradykinin and A-23187. In the presence of
NG-nitro-L-arginine methyl ester,
TNF- produced no further inhibition on the vasorelaxation induced by
these vasodilators. With the use of 4,5-diaminofluorescein diacetate
fluorescence imaging analysis, bradykinin was found to increase nitric
oxide (NO) concentrations in the endothelium of isolated bovine small
coronary arteries, which was inhibited by TNF-. Pretreatment of the
arteries with desipramine (10 µM), an inhibitor of acidic
sphingomyelinase, tiron (1 mM), a superoxide scavenger, and
polyethylene glycol-superoxide dismutase (100 U/ml) largely restored
the inhibitory effect of TNF- on bradykinin- and A-23187-induced
vasorelaxation. In addition, TNF- activated acidic sphingomyelinase
and increased ceramide levels in coronary endothelial cells. We
conclude that TNF- inhibits NO-mediated endothelium-dependent
vasorelaxation in small coronary arteries via sphingomyelinase
activation and consequent superoxide production in endothelial cells.
tumor necrosis factor; lipids; nitric oxide; free radicals
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