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From the Houston VA Medical Center (W.D., M.H.K., N.C., K.J.P., A.I.S.) and the Departments of Medicine (W.D., M.H.K., N.C., A.I.S.) and Pharmacology (W.D.), Baylor College of Medicine, Houston, Tex.
Correspondence to Dr William Durante, Houston VA Medical Center, Building 109, Room 116, 2002 Holcombe Blvd, Houston, TX 77030.
Abstract Since recent studies demonstrate that vascular
smooth muscle cells synthesize two distinct guanylate
cyclasestimulatory gases, NO and CO, we examined possible regulatory
interactions between these two signaling molecules. Treatment of rat
aortic smooth muscle cells with the NO donors, sodium nitroprusside,
S-nitroso-N-acetyl-penicillamine, or
3-morpholinosydnonimine, increased heme oxygenase-1 (HO-1) mRNA and
protein levels in a concentration- and time-dependent manner. Both
actinomycin D and cycloheximide blocked NO-stimulated HO-1 mRNA and
protein expression. Nuclear run-on experiments demonstrated that NO
donors increased HO-1 gene transcription between 3- and 6-fold. In
contrast, NO donors had no effect on the stability of HO-1 mRNA.
Incubation of vascular smooth muscle cells with the membrane-permeable
cGMP analogues, dibutyryl cGMP and 8-bromo-cGMP, failed to induce HO-1
gene expression. Treatment of vascular smooth muscle cells with NO
donors also stimulated the production and release of CO, as
demonstrated by the CO-dependent increase in intracellular cGMP levels
in coincubated platelets. Finally, incubating vascular smooth muscle
cells with interleukin-1ß and tumor necrosis factor-
induced NO
synthesis and also significantly increased the level of HO-1 protein.
The cytokine-stimulated production of both NO and HO-1 protein in
smooth muscle cells was blocked by the NO synthase inhibitor
methyl-L-arginine. These results demonstrate that
exogenously administered or endogenously released NO stimulates HO-1
gene expression and CO production in vascular smooth muscle cells. The
ability of NO to induce HO-catalyzed CO release from vascular smooth
muscle cells provides a novel mechanism by which NO might modulate
soluble guanylate cyclase and, thereby, vascular smooth muscle cell and
platelet function.
Key Words: nitric oxide carbon monoxide heme oxygenase
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