HMG-CoA reductase inhibitor enhances inducible nitric oxide synthase expression in rat vascular smooth muscle cells; involvement of the Rho/Rho kinase pathway

Abstract

Little is known about the mechanism by which HMG-CoA reductase inhibitors affect inducible nitric oxide synthase (iNOS) expression. We investigated the effect of HMG-CoA reductase inhibitor cerivastatin on iNOS expression in cultured rat vascular smooth muscle cells (VSMCs). Quiescent VSMCs were incubated with or without various concentrations of drugs as follows: cerivastatin, C3 exoenzyme or Y-27632. Then, pretreated VSMCs were stimulated by a vehicle or interleukin (IL)-1β (10 ng/ml). Treatment of VSMCs with cerivastatin (10 −7–10 −5 mol/l), which inhibits isoprenylation of Rho and other small G proteins, significantly increased nitrite/nitrate (NOx) production and upregulated the expression of iNOS mRNA in IL-1β-stimulated VSMCs. This effect of cerivastatin was abolished by cotreatment with mevalonate (2×10 −4 mol/l) or geranylgeranyl-pyrophosphate (GGPP) (10 −5 mol/l), but not by farnesyl-pyrophosphate (10 −5 mol/l). Furthermore, C3 exoenzyme (50 μg/ml), an inactivator of Rho protein, and Rho kinase inhibitor Y-27632 (10 −5 mol/l) also enhanced NOx production and the expression of iNOS mRNA in IL-1β-stimulated VSMCs. Immunocytochemical study revealed that cerivastatin, C3 exoenzyme and Y-27632 did not affect the nuclear translocation of nuclear factor-κB in IL-1β-stimulated VSMCs. Our study suggests that cerivastatin stimulates iNOS expression in IL-1β treated VSMCs by its inhibitory effect on Rho/Rho kinase pathway. In addition, this effect of cerivastatin, by enhancing iNOS expression, may contribute to the prevention of restenosis after percutaneous coronary intervention and protect against atherothrombosis.