Nitrite and nitroglycerin induce rapid release of the vasodilator ATP from erythrocytes: Relevance to the chemical physiology of local vasodilation

Extracellular ATP released from circulating erythrocytes induces vasodilation by stimulating receptor-mediated endothelium NO/EDRF (endothelium-derived relaxing factor) production. We report that pre-stimulation of freshly isolated human erythrocytes with physiological nitrite (100 nM NO2-) or pharmacological nitroglycerin (10 μM) concentrations resulted in >200% spike in ATP release, which was detected on resuspending the cells in fresh medium. The observed response was instantaneous following NO2- pre-stimulation but a delay of ∼20 s followed nitroglycerin pre-stimulation, reflecting the time required for prodrug activation within the erythrocyte to its vasoactive metabolites, NO2- and NO. The data provided here are consistent with ATP being a conveyor of a NO-induced vasodilatory signal from the erythrocyte to the endothelium. Extended erythrocyte pre-stimulation with the NO donors resulted in a dose-dependent decrease in extracellular ATP, which would attenuate the signal in intact vessels to prevent excessive vasodilation. Importantly, our study constitutes the first report of enhanced vasodilator (ATP) release following human erythrocyte pre-stimulation by an endogenous (NO2-) or pharmacological (nitroglycerin) NO donor. The relevance of our findings to the therapeutic effects of nitroglycerin as well as to nitrate tolerance is discussed.