Antithrombotic effects of peroxynitrite: inhibition and reversal of aggregation in human platelets

The inhibition of platelet aggregation by peroxynitrite, a reactive oxygen species derived from the interaction of nitric oxide (NO) and superoxide, was examined in platelet-rich plasma. In this report, we have used a preparation of peroxynitrite that was free of H2O2 and MnO2. As such, peroxynitrite dose-dependently (50-200 microM) inhibited aggregation of human platelets stimulated by ADP (5 microM), collagen (0.5 microgram), thrombin (0.5U/microL) and U46619 (1 microM). In addition, peroxynitrite reversed platelet aggregation induced by collagen, ADP, and thrombin. Peroxynitrite, preincubated with platelet-poor plasma or albumin (7%) for 30 min, did not alter the inhibition of platelet aggregation. This suggested that the inhibitory action of peroxynitrite may be due to nitrosylation of proteins, which by themselves possess activity, rather than conversion to NO or NO donors. Furthermore, we show that peroxynitrite increased the cGMP level only at 200 microM concentrations, further suggesting that the action of peroxynitrite was not completely due to its conversion to NO or NO donors.