Inhibition of prostaglandin H synthase and activation of 12-lipoxygenase by 8,11,14,17-eicosatetraenoic acid in human endothelial cells and platelets

The effects of the marine fatty acid 20:4n-3, an isomer of arachidonic acid (20:4n-6), have been compared to that of 20:5n-3 on 20:4n-6 oxygenation in human platelets and endothelial cells. In platelets, 20:4n-3 added along w ith 20:4n-6 was as potent as 20:5n-3 in inhibiting prostaglandin H synthase (PGH synthase) activity. From 2.5- to 10 mu M of 20:4n-6, the synthesis of thromboxane Bz and 12-hydroxy-5,8,10-heptadecatrienoic acid, reflecting th e PGH/thromboxane synthase activity, was lowered by 5 and 10 mu M Of both f atty acids. In contrast, 20:4n-3, but not 20:5n-3, strongly stimulated the lipoxygenase activity at each concentration of 20:4n-6 used whatever the am ount of 20:4n-3 added. The effects of both n-3 polyunsaturated fatty acids on endothelial cell PGH/prostacyclin synthases were compared after 2- and 2 4-hr incubation with the cells, leading to moderate (2 hr) and high (24 hr) concentrations of these fatty acids in membrane phospholipids. The incorpo ration of 20:4n-3 and 20:5n-3 occurred mostly in phosphatidylcholine and ph osphatidylethanolamine and did not after the 20:4n-6 lever of phospholipid classes after 2-hr supplementation, whereas it was drastically decreased af ter 24 hr. The synthesis of prostacyclin obtained after cell stimulation by 0.1 U/mL thrombin was unaffected by the fatty acid modifications induced a fter 2-hr supplementation whereas it was strongly depressed after 24 hr. It was concluded that 20:4n-3 is not an agonist for platelet activation, desp ite its close structural analogy with 20:4n-6, and is as potent as 20:5n-3 in inhibiting PGH synthase activities, showing that the double bond at C5 i s not necessary for inhibition. In contrast, the oxygenation of 20:4n-6 by 12-lipoxygenase was stimulated by 20:4n-3 but not by 20:5n-3, which might b e related to the efficient oxygenation of 20:4n-3 by this enzyme compared w ith 20:5n-3. (C) 1999 Elsevier Science Inc.