Study of the two pathways for arachidonate oxygenation in blood platelets.
During collagen-induced blood platelet aggregation, arachidonic acid is set free from membrane phospholipids and subsequently converted into 12-hydroxyeicosatetraenoic acid by arachidonate lipoxygenase and into thromboxane A2, 12-hydroxyheptadecatrienoic acid (HETE) and malondialdehyde by cyclooxygenase and thromboxane synthase. Lipoxygenase and cyclooxygenase have optimal activity at neutral to basic pH, while the thromboxane synthase is pH-independent between 5 and 9. These enzymes are membrane-bound. The cyclooxygenase is rapidly inactivated upon membrane disruption by nonionic detergents or phospholipid degradation with phospholipase A2. It was found that platelet phospholipase A2 preferentially splits off fatty acid with four double bonds. Eicosatetraynoic acid was used to investigate the physiological function of the arachidonate lipoxygenase during collagen-induced aggregation of rat blood platelets. This fatty acid is a more efficient inhibitor of lipoxygenase than of cyclooxygenase. At an inhibitor concentration of 0.6 microgram/ml, platelet aggreation, 12-hydroxyeicosatetraenoic acid production as well as 15-hydroxytryptamine release are completely inhibited, while there is an apparent stimulation of the cyclooxygenase. These results indicate that arachidonate lipoxygenase is essential for irreversible blood platelet aggregation.
D000818:Animals; D001095:Arachidonic Acids / Q000097:blood* / Q000378:metabolism / Q000494:pharmacology; D001792:Blood Platelets / Q000378:metabolism*; D002417:Cattle; D003094:Collagen; D006863:Hydrogen-Ion Concentration; D006880:Hydroxy Acids / Q000378:metabolism; D008084:Lipoxygenase / Q000097:blood; D010974:Platelet Aggregation / Q000187:drug effects; D011092:Polyethylene Glycols; D011449:Prostaglandin Endoperoxides / Q000494:pharmacology; D011451:Prostaglandin-Endoperoxide Synthases / Q000097:blood; D051381:Rats; D013928:Thromboxane A2 / Q000378:metabolism; D013929:Thromboxane B2 / Q000378:metabolism
D001095:Arachidonic Acids; D006880:Hydroxy Acids; D011449:Prostaglandin Endoperoxides; D011092:Polyethylene Glycols; D013929:Thromboxane B2; D013928:Thromboxane A2; D003094:Collagen; D008084:Lipoxygenase; D011451:Prostaglandin-Endoperoxide Synthases