Increased thromboxane B2 biosynthesis in platelets.
The synthesis of thromboxane B2 is increased in platelets from rabbits with experimental hypercholesterolemia, but the increase is not due to increased phospholipids hydrolysis. We have clarified the mechanism for the increased thromboxane synthesis. The biosyntheses of prostaglandin H2 and thromboxane B2 were unaffected by superoxide dismutase, xanthine oxidase, mannitol, or benzoate in other experiments designed to study the possible involvement of reactive oxygen species. These results suggest that O2.- and OH were not likely to be involved as intermediates in the synthesis of prostaglandin H2 and thromboxane B2 in platelets. The rate of prostaglandin H2 biosynthesis was promoted in deuterium oxide, and this deuterium oxide enhancement effect was reversed by 2,5-diphenylfuran, suggesting that singlet oxygen may be involved in prostaglandin H2 biosynthesis. The biosynthesis of prostaglandin H2 was promoted by ADP-Fe3+ but inhibited by EDTA and EDTA-Fe3+. The effect of ADP-Fe3+ could not be replaced by EDTA-Fe3+. The effects of glutathione, glutathione peroxidase and H2O2 on cyclooxygenase and thromboxane synthetase were studied by using partially purified enzymes and platelet microsomes. Glutathione and glutathione peroxidase inhibited the activity of cyclooxygenase but did not inhibit that of thromboxane synthetase. H2O2 caused the inactivation of cyclooxygenase, but the addition of H2O2 did not inhibit the formation of thromboxane B2 from prostaglandin H2. An examination of glutathione concentration and glutathione peroxidase activity in platelets from normal and experimentally hypercholesterolemic rabbits demonstrated that both were decreased in platelets from later group. The observed alterations in glutathione levels and glutathione peroxidase activity are large enough to cause increased thromboxane B2 synthesis in platelets but the possibility that other unidentified factors may also contribute cannot be excluded.
D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000097:blood; D001792:Blood Platelets / Q000378:metabolism*; D002374:Catalase / Q000097:blood; D005290:Ferric Compounds / Q000494:pharmacology; D005979:Glutathione Peroxidase / Q000097:blood; D006937:Hypercholesterolemia / Q000097:blood*; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology; D009251:NADPH-Ferrihemoprotein Reductase / Q000097:blood; D011450:Prostaglandin Endoperoxides, Synthetic / Q000096:biosynthesis; D044262:Prostaglandin H2; D011463:Prostaglandins H / Q000096:biosynthesis; D011817:Rabbits; D013482:Superoxide Dismutase / Q000097:blood; D013929:Thromboxane B2 / Q000096:biosynthesis*; D013931:Thromboxanes / Q000096:biosynthesis*
D001095:Arachidonic Acids; D005290:Ferric Compounds; D011450:Prostaglandin Endoperoxides, Synthetic; D011463:Prostaglandins H; D013931:Thromboxanes; D016718:Arachidonic Acid; D044262:Prostaglandin H2; D013929:Thromboxane B2; D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D009251:NADPH-Ferrihemoprotein Reductase