Stimulation of Platelet Nitric Oxide Production by Nebivolol Prevents Thrombosis

Abstract

Objective— dl -Nebivolol, a selective β1-adrenergic receptor antagonist, besides its hypotensive activity exerts vasodilatory and platelet inhibitory effects in vitro by a mechanism involving nitric oxide (NO). Our aim was to evaluate whether nebivolol exerts in vivo antithrombotic effects, to unravel the mechanism of this action and to clarify the relative roles of its 2 enantiomers: d - and l -nebivolol. Methods and Results— In wild-type mice, dl -nebivolol, l -nebivolol, and d -nebivolol, but not bisoprolol, reduced mortality consequent to platelet pulmonary thromboembolism induced by the intravenous injection of collagen plus epinephrine (−44%, −45%, −29%, respectively; P <0.05), whereas in eNOS −/− mice only dl -nebivolol and d -nebivolol were effective. dl -Nebivolol, l - and d -nebivolol reduced photochemical damage-induced femoral artery thrombosis in wild-type mice, whereas in eNOS −/− mice only dl -nebivolol and d -nebivolol were active. Moreover, dl -nebivolol and l -nebivolol increased plasma, urinary-, and platelet-derived nitrites and nitrates (NOx), NO degradation products, in wild-type but not in eNOS −/− mice. In vivo platelet activation, assessed by platelet P-selectin expression, was reduced by dl -nebivolol and l - and d -nebivolol in wild-type mice but only by dl -nebivolol and d -nebivolol in eNOS −/− mice. In bone marrow–transplanted, chimeric mice with only blood cells, and not the endothelium, producing NO dl -nebivolol and l -nebivolol maintained their antithrombotic activity, whereas they lose it in chimeras with only endothelium, and not blood cells, producing NO. In vitro, with isolated platelets, dl -nebivolol and l -nebivolol, but not d -nebivolol and bisoprolol, increased platelet cGMP and NOx formation. Treatment with dl -nebivolol and l -nebivolol increased phophorylated eNOS in platelets. Conclusions— Our data show that dl -nebivolol exerts an antithrombotic activity by stimulating the formation of NO by platelets, and that this effect is generated by its l -enantiomer, whereas the d -enantiomer exerts a weak antiplatelet effect because of β−adrenergic receptor–independent stimulation of adenyly cyclase. These results confirm that platelet-derived NO plays a role in thrombosis prevention and it may represent a target of pharmacological intervention.