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1 Department of Physiology, Wayne State University School of Medicine, Detroit, Michigan 48201; and 2 Department of Medicine and Cell Biology, State University of New York Health Sciences Center and Veterans Affairs New York Harbor Healthcare Center, Brooklyn, New York 11203
Inducible nitric oxide synthase (iNOS) in vascular smooth
muscle cells (VSMCs) is upregulated in arterial injury and plays a role
in regulating VSMC proliferation and restenosis. Inflammatory cytokines
[e.g., interleukin-1
(IL-1)] released during vascular injury induce iNOS. Small GTP-binding proteins of the Ras superfamily play a major role in IL-1-dependent signaling pathways.
In this study, we examined the role of Rho GTPases in regulating iNOS expression in VSMCs. Treatment of VSMCs with mevastatin, which inhibits
isoprenylation of Rho and other small GTP-binding proteins, produced
significantly higher amounts of IL-1-evoked NO and iNOS protein
compared with control. Similarly, bacterial toxins [Toxin B from
Clostridium difficile and C3 ADP-ribosyl transferase (C3) toxin
from Clostridium botulinium] that specifically inactivate Rho proteins increased NOS products (NO and citrulline) and iNOS expression. Toxin B increased the activity of iNOS promoter-reporter construct in VSMCs. Both toxins enhanced IL-1-stimulated iNOS expression and NO production. These data demonstrate for the first time
that inhibition of Rho induces iNOS and suggest a role for Rho protein
in IL-1-stimulated NO production in VSMCs.
nitric oxide synthase; inducible; guanosine 5'-triphosphate phosphohydrolase
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