Intraplatelet tetrahydrobiopterin plays an important role in regulating canine coronary arterial thrombosis by modulating intraplatelet nitric oxide and superoxide generation

Background-Platelet-derived nitric oxide inhibits platelet aggregation via constitutive NO synthase (NOS). Tetrahydrobiopterin (BH4), a cofactor of NOS, augments NO formation, whereas its deficiency decreases NO bioactivity and increases superoxide generation by NOS. The roles of intraplatelet BH4 in platelet aggregation and thrombus formation, however, are unknown. Accordingly, we investigated whether intraplatelet BH4 is involved in regulating cyclic flow variations (CFVs) and platelet aggregation in a canine model with stenosed and endothelium-injured coronary arteries that mimics acute coronary syndromes in humans. Methods and Results-After developing CFVs, dogs received saline or BH4 (10 or 30 mg/kg) intravenously. Intraplatelet BH4 and cGMP levels were decreased and intraplatelet nitrotyrosine production was increased during CFVs. ADP- and U46619-induced ex vivo platelet aggregation and platelet P-selectin expression were augmented during CFVs. BH4 administration restored intraplatelet BH4 and cGMP levels and decreased intraplatelet nitrotyrosine production, resulting in reduced CFVs and inhibited ex vivo platelet aggregation and platelet P-sclectin expression. CFVs again developed after N-G-monomethyl-L-arginine, an inhibitor of NOS, in BH4-treated dogs. Ex vivo platelet NOS activity at baseline, during CFVs, and after BH, administration did not differ. Conclusions-Intraplatelet BH4 may play an important role in regulating thrombus formation by modulating platelet-derived nitric oxide and superoxide generation by platelet NOS.