Hydrogen peroxide and methyl mercury are primary stimuli of eicosanoid release in human platelets.
Hydrogen peroxide (H2O2) and methyl mercury induced the liberation of arachidonate and its metabolites from human washed platelets. [14C]Eicosanoids were extracted from the supernatants of [14C] arachidonate-prelabelled platelets and analysed by thin layer chromatography and radioscanning. Thromboxane B2 (TXB2), 12(S)-hydroxy-5,8,10-heptadecatrienoic acid (HHT) and 12(S)-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE) were found as stable metabolites, together with unreacted arachidonate. In the presence of dazoxiben, a shift in eicosanoid metabolism was observed towards prostaglandin E2 (PGE2), prostaglandin D2 (PGD2) and prostaglandin F2 alpha (PGF2 alpha), while in the presence of indomethacin there was a shift towards 12-HETE and unmetabolized arachidonate. The concentration pattern of those metabolites resembled that found with the physiological agonist, thrombin. H2O2 and methyl mercury also induced platelet shape change, aggregation and secretion. The EC50 values for the induction of shape change and aggregation were 27 and 850 mumol/l for H2O2 and 0.33 and 2.7 mumol/l for methyl mercury, respectively. The [3H]serotonin release required higher stimulus concentrations and amounted to 45% with 2 mumol/l H2O2 and to 16% with 3 mumol/l methyl mercury. These effects on platelet function were absent in platelets exposed to acetylsalicylic acid and prevented by indomethacin, the prostaglandin H2 (PGH2)/thromboxane A2 (TXA2) receptor antagonist, daltroban, and the functional antagonist, iloprost. In contrast, none of these drugs suppressed the formation of [14C]eicosanoids, indicating that the platelet activation by H2O2 and methyl mercury essentially requires previous PGH2/TXA2 formation. As expected, the thromboxane synthase inhibitor, dazoxiben, did not prevent, but instead potentiated the activation by H2O2 and methyl mercury through accumulated PGH2.(ABSTRACT TRUNCATED AT 250 WORDS)
Hornberger
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W
Institut für Klinische Chemie, Klinikum Mannheim der Universität Heidelberg, F.R.G.
Patscheke
H
H
D001095:Arachidonic Acids / Q000378:metabolism; D001241:Aspirin / Q000494:pharmacology; D001792:Blood Platelets / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015777:Eicosanoids / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D011464:Epoprostenol / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D016285:Iloprost; D007093:Imidazoles / Q000494:pharmacology; D007213:Indomethacin / Q000494:pharmacology; D008767:Methylmercury Compounds / Q000494:pharmacology*; D010974:Platelet Aggregation / Q000187:drug effects; D012701:Serotonin / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology
D001095:Arachidonic Acids; D015777:Eicosanoids; D007093:Imidazoles; D008767:Methylmercury Compounds; C029166:dazoxiben; D012701:Serotonin; D006861:Hydrogen Peroxide; D011464:Epoprostenol; D013917:Thrombin; D016285:Iloprost; D001241:Aspirin; C004925:methylmercuric chloride; D007213:Indomethacin