Tetrahydrobiopterin Deficiency and Nitric Oxide Synthase Uncoupling Contribute to Atherosclerosis Induced by Disturbed Flow

OBJECTIVE: Tetrahydrobiopterin (BH(4)) is a critical cofactor for nitric oxide (NO) synthesis by NO synthase (NOS). Recently, we demonstrated that disturbed flow produced by partial carotid ligation decreases BH(4) levels in vivo. We therefore aimed to determine whether atherosclerosis induced by disturbed flow is due to BH(4) deficiency and NOS uncoupling and whether increasing BH(4) would prevent endothelial dysfunction, plaque inflammation, and atherosclerosis. METHODS AND RESULTS: We produced a region of disturbed flow in apolipoprotein E(-/-) mice using partial carotid ligation and fed these animals a high-fat diet. This caused endothelial NOS uncoupling as characterized by increased vascular superoxide production, altered vascular reactivity, and a change in endothelial NOS migration on low-temperature gel. These perturbations were accompanied by severe atherosclerosis, infiltration of T cells and macrophages, and an increase in cytokine production. Treatment with BH(4) recoupled NOS, decreased superoxide production, improved endothelium-dependent vasodilatation, and virtually eliminated atherosclerosis. BH(4) treatment also markedly reduced vascular inflammation and improved the cytokine milieu induced by disturbed flow. CONCLUSIONS: Our results highlight a key role of BH(4) deficiency and NOS uncoupling in atherosclerosis induced by disturbed flow and provide insight into the effect of modulating vascular BH(4) levels on atherosclerosis and inflammation at these sites of the circulation.