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NADPH oxidases (NOXs) contribute to platelet activation by a largely unknown mechanism. Here, we studied the effect of the novel NOX inhibitor 2‐acetylphenothiazine (2‐APT) on human platelet functional responses and intracellular signaling pathways.

The generation of superoxide ions was assessed by single cell imaging on adhering platelets using dihydroethidium (DHE), while other reactive oxygen species (ROS) were detected with 5‐(and‐6)‐carboxy‐2′,7′‐dichlorodihydrofluorescein diacetate (CM‐H₂‐DCFDA). Whole blood thrombus formation, washed platelet aggregation, integrin αIIbβ3 inside‐out signalling, Syk phosphorylation and PKC activation were analysed to understand the functional consequences of NOX inhibition by 2‐APT in platelets.

Superoxide ion generation stimulated by platelet adhesion on collagen and fibrinogen …