Objective-Platelets undergoing anoxia-reoxygenation (AR) simultaneously increase reactive oxygen species (ROS) and thromboxane (Tx) B 2 . Our aim was to assess whether there is an interplay between activation of NOX2, the catalytic subunit of NADPH oxidase, and platelet TxB 2 in vitro and in vivo. Methods and Results-Platelets that underwent AR had enhanced ROS. This was associated with NOX2 activation and was inhibited by incubation with NOX2-blocking peptide. AR was associated with TxB 2 and isoprostane production, which were inhibited by NOX2-blocking peptide, vitamin C, and the inhibitor of phospholipase A 2 . Platelet incubation with 100 mol/L aspirin fully prevented AR-induced TxA 2 but did not affect isoprostane production. We included 56 aspirin-treated patients undergoing elective percutaneous coronary intervention (PCI) who were randomly allocated to receive either placebo or intravenous infusion of 1 g of vitamin C. Blood TxB 2 , isoprostanes, and soluble NOX2-derived peptide, a marker of systemic NADPH oxidase activation, significantly increased at 60 and 120 minutes after PCI in placebo-treated but not in vitamin C-treated patients. Conclusion-AR is associated with overproduction of platelet TxB 2 and isoprostanes, which is dependent on NOX2-dependent ROS generation. Low doses of aspirin are unable to prevent TxB 2 formation in patients who undergo PCI. Key Words: aspirin Ⅲ thromboxanes Ⅲ NOX2 Ⅲ vitamin C Ⅲ isoprostanes E xperimental models of ischemia-reperfusion are associated with a burst of reactive oxygen species (ROS), which may have deleterious effect in the coronary circulation. 1,2 Accordingly, percutaneous coronary intervention (PCI), a typical in vivo model of ischemia-reperfusion, has been associated with overproduction of isoprostanes, which are markers of oxidative stress, as well as with impaired microcirculatory flow. 3 Furthermore, in vitro experiments of anoxia-reoxygenation (AR) demonstrated enhanced platelet activation via formation of thromboxane (Tx) A 2 ; this may negatively influence the circulatory flow by predisposing to thrombotic complication. 4 AR-induced platelet TxB 2 formation follows arachidonic acid release by platelet membrane via ROS-induced phospholipase A 2 (PLA 2 ) activation. 5 However, the mechanism accounting for ROS formation in platelets undergoing AR is still undefined.NADPH oxidase, the most important cellular producer of ROS, is present not only in the immune innate cells but also in vessel walls and platelets. 6 Platelets express NOX2, the catalytic subunit of NADPH oxidase, along with the other cytosolic subunits. 7,8 Genetic deficiency of NOX2 is associated with low/absent platelet ROS formation and platelet dysfunction, thus suggesting that NOX2 activation is implicated in platelet activation and ROS formation. 9 We hypothesized that AR could enhance platelet TxB 2 formation via NOX2 activation and that antioxidants are able to prevent it. To this end, we analyzed in vitro whether NOX2 plays a role in the ROS formation elicited by AR and whether the...