Withdrawal of 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase Inhibitors Elicits Oxidative Stress and Induces Endothelial Dysfunction in Mice

Abstract

3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) improve endothelial function. We determined whether withdrawal of statin therapy affects endothelium-dependent relaxation in mice and studied the underlying mechanism. Mice were treated with daily injections of cerivastatin (2 mg/kg per day SC), atorvastatin (1 and 10 mg/kg per day SC), or placebo. Vascular reactivity was studied in aortic rings from these mice after 10 days of treatment and after cessation of therapy for several days. Both statins improved endothelium-dependent relaxation to acetylcholine. Compared with control, withdrawal of statin treatment transiently (from day 4 to 7) attenuated endothelium-dependent relaxation. In vessels from animals subjected to atorvastatin withdrawal, the antioxidant tiron restored relaxations. Vascular superoxide anion generation was unaffected by statin therapy but was increased during withdrawal. In mice lacking the gp91phox subunit of the NADPH oxidase, no attenuation of acetylcholine-induced relaxation and no increase in superoxide generation were observed after withdrawal of atorvastatin. In human umbilical vein endothelial cells, statins, which decrease the membrane association of NADPH oxidase–activating Rac-1, increased the activity of this GTPase in whole-cell lysates. Withdrawal of statins induced a translocation of Rac-1 from the cytosol to the membrane and transiently increased NADPH-induced lucigenin chemiluminescence in membrane preparations. Rac-1 inactivation by Clostridium difficile toxin B inhibited the cerivastatin-induced oxygen radical production in human umbilical vein endothelial cells. These observations indicate that the withdrawal of statins induces endothelial dysfunction. The underlying mechanism involves activation of a gp91phox-containing NADPH oxidase by Rac-1 and the subsequent scavenging of endothelium-derived NO by superoxide anions generated from this enzyme.