Hypertension, Vol 22, 34-39, Copyright © 1993 by American Heart Association
K Kanno, Y Hirata, T Imai and F Marumo
To elucidate whether cytokines induce nitric oxide synthase in vascular
smooth muscle cells, we studied the effects of human recombinant
interleukin-1 beta on the synthesis and release of nitric oxide in cultured
rat vascular smooth muscle cells by measurement of NO2-/NO3- levels.
Furthermore, we performed Northern blot analysis using subcloned polymerase
chain reaction products as probes for constitutive and inducible nitric
oxide synthase. Interleukin-1 beta dose dependently (1 to 20 ng/mL)
stimulated NO2-/NO3- production as a function of time. Northern blotting
demonstrated the interleukin-1 beta- induced expression of messenger RNA
for an inducible but not for the constitutive nitric oxide synthase after 3
hours. NG-Monomethyl L- arginine completely blocked the interleukin-1
beta-induced NO2-/NO3- production, the effect of which was reversed by
L-arginine but not by D- arginine. Dexamethasone inhibited the
interleukin-1 beta-induced NO2- /NO3- production in a dose-dependent manner
(10(-9) to 10(-7) M) and the interleukin-1 beta-inducible nitric oxide
synthase messenger RNA levels. Neither a calmodulin inhibitor (W-7) nor a
protein kinase C inhibitor (staurosporine) showed any effects on the
induction of nitric oxide synthase transcripts or production of NO2-/NO3-
stimulated by interleukin-1 beta, whereas cycloheximide and actinomycin D
completely inhibited the basal and stimulated NO2-/NO3- production. These
data demonstrate for the first time that interleukin-1 beta induces gene
expression of inducible nitric oxide synthase and its de novo protein
synthesis in rat vascular smooth muscle cells, thereby leading to
generation of nitric oxide via Ca2+/calmodulin-independent and protein
kinase C-independent mechanisms.
ARTICLES
Induction of nitric oxide synthase gene by interleukin in vascular smooth muscle cells
Second Department of Internal Medicine, Tokyo Medical and Dental University, Japan.
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