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. 1992 Jun;106(2):267–274. doi: 10.1111/j.1476-5381.1992.tb14327.x

Biological activity and metabolism of 20-hydroxyeicosatetraenoic acid in the human platelet.

E Hill 1, F Fitzpatrick 1, R C Murphy 1
PMCID: PMC1907511  PMID: 1327375

Abstract

1. The cytochrome P-450 metabolite of arachidonic acid, 20-hydroxyeicosatetraenoic acid (20-HETE), was found to be a potent, dose-dependent inhibitor of platelet aggregation and inhibitor of thromboxane biosynthesis induced by arachidonic acid (IC50 5.2 +/- 1.5 microM), A23187 (IC50 16.2 +/- 5.4 microM), and U46619 (IC50 7.8 +/- 2.4 microM). 20-HETE did not inhibit thrombin-induced aggregation. 2. The human platelet metabolized 20-HETE to a series of novel metabolites formed by cyclo-oxygenase as well as lipoxygenase pathways. The structures of the metabolites were identified by mass spectrometry as 20-hydroxy-thromboxane B2, 12,17-dihydroxyheptadecatrienoic acid, 12,20-dihydroxyeicosatetraenoic acid, and 11,20-dihydroxyeicosatetraenoic acid. 3. The identification of the 11-hydroxy metabolite of 20-HETE suggests that 20-HETE is less efficiently cyclized to an endoperoxide intermediate by cyclo-oxygenase than is arachidonate. 4. Although some biological activity of 20-HETE may be related to competition with endogenous arachidonate for cyclo-oxygenase metabolism, the predominant mechanism of action of 20-HETE appears to be through antagonism of the prostaglandin H2/thromboxane A2 receptor.

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Selected References

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  1. Abraham N. G., Feldman E., Falck J. R., Lutton J. D., Schwartzman M. L. Modulation of erythropoiesis by novel human bone marrow cytochrome P450-dependent metabolites of arachidonic acid. Blood. 1991 Sep 15;78(6):1461–1466. [PubMed] [Google Scholar]
  2. Aharony D., Smith J. B., Silver M. J. Regulation of arachidonate-induced platelet aggregation by the lipoxygenase product, 12-hydroperoxyeicosatetraenoic acid. Biochim Biophys Acta. 1982 Oct 8;718(2):193–200. doi: 10.1016/0304-4165(82)90219-7. [DOI] [PubMed] [Google Scholar]
  3. Bailey J. M., Bryant R. W., Whiting J., Salata K. Characterization of 11-HETE and 15-HETE, together with prostacyclin, as major products of the cyclooxygenase pathway in cultured rat aorta smooth muscle cells. J Lipid Res. 1983 Nov;24(11):1419–1428. [PubMed] [Google Scholar]
  4. Bernstrom K., Malcolm K., McGee J., Maclouf J., Levy-Toledano S., Falck J. R., Fitzpatrick F. A. 14, 15-cis-episulfide-eicosatrienoic acid, an 'epoxygenase' eicosanoid analog, inhibits ionophore- but not thrombin-induced platelet aggregation. Mol Pharmacol. 1991 Feb;39(2):114–119. [PubMed] [Google Scholar]
  5. Brüne B., Ullrich V. 12-hydroperoxyeicosatetraenoic acid inhibits main platelet functions by activation of soluble guanylate cyclase. Mol Pharmacol. 1991 May;39(5):671–678. [PubMed] [Google Scholar]
  6. Capdevila J. H., Falck J. R., Dishman E., Karara A. Cytochrome P-450 arachidonate oxygenase. Methods Enzymol. 1990;187:385–394. doi: 10.1016/0076-6879(90)87045-5. [DOI] [PubMed] [Google Scholar]
  7. Carroll M. A., Sala A., Dunn C. E., McGiff J. C., Murphy R. C. Structural identification of cytochrome P450-dependent arachidonate metabolites formed by rabbit medullary thick ascending limb cells. J Biol Chem. 1991 Jul 5;266(19):12306–12312. [PubMed] [Google Scholar]
  8. Escalante B., Erlij D., Falck J. R., McGiff J. C. Effect of cytochrome P450 arachidonate metabolites on ion transport in rabbit kidney loop of Henle. Science. 1991 Feb 15;251(4995):799–802. doi: 10.1126/science.1846705. [DOI] [PubMed] [Google Scholar]
  9. Escalante B., Sessa W. C., Falck J. R., Yadagiri P., Schwartzman M. L. Vasoactivity of 20-hydroxyeicosatetraenoic acid is dependent on metabolism by cyclooxygenase. J Pharmacol Exp Ther. 1989 Jan;248(1):229–232. [PubMed] [Google Scholar]
  10. Fitzpatrick F. A., Murphy R. C. Cytochrome P-450 metabolism of arachidonic acid: formation and biological actions of "epoxygenase"-derived eicosanoids. Pharmacol Rev. 1988 Dec;40(4):229–241. [PubMed] [Google Scholar]
  11. Gilman A. G. A protein binding assay for adenosine 3':5'-cyclic monophosphate. Proc Natl Acad Sci U S A. 1970 Sep;67(1):305–312. doi: 10.1073/pnas.67.1.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hamberg M., Samuelsson B. On the mechanism of the biosynthesis of prostaglandins E-1 and F-1-alpha. J Biol Chem. 1967 Nov 25;242(22):5336–5343. [PubMed] [Google Scholar]
  13. Hamberg M., Samuelsson B. Prostaglandin endoperoxides. Novel transformations of arachidonic acid in human platelets. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3400–3404. doi: 10.1073/pnas.71.9.3400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hatzelmann A., Ullrich V. The omega-hydroxylation of arachidonic acid by human polymorphonuclear leukocytes. Eur J Biochem. 1988 Apr 15;173(2):445–452. doi: 10.1111/j.1432-1033.1988.tb14019.x. [DOI] [PubMed] [Google Scholar]
  15. Hemler M. E., Crawford C. G., Lands W. E. Lipoxygenation activity of purified prostaglandin-forming cyclooxygenase. Biochemistry. 1978 May 2;17(9):1772–1779. doi: 10.1021/bi00602a031. [DOI] [PubMed] [Google Scholar]
  16. Levy-Toledano S., Maclouf J., Bryon P., Savariau E., Hardisty R. M., Caen J. P. Human platelet activation in the absence of aggregation: a calcium-dependent phenomenon independent of thromboxane formation. Blood. 1982 May;59(5):1078–1085. [PubMed] [Google Scholar]
  17. Maclouf J. A., Murphy R. C. Transcellular metabolism of neutrophil-derived leukotriene A4 by human platelets. A potential cellular source of leukotriene C4. J Biol Chem. 1988 Jan 5;263(1):174–181. [PubMed] [Google Scholar]
  18. Marcus A. J., Safier L. B., Ullman H. L., Broekman M. J., Islam N., Oglesby T. D., Gorman R. R. 12S,20-dihydroxyicosatetraenoic acid: a new icosanoid synthesized by neutrophils from 12S-hydroxyicosatetraenoic acid produced by thrombin- or collagen-stimulated platelets. Proc Natl Acad Sci U S A. 1984 Feb;81(3):903–907. doi: 10.1073/pnas.81.3.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Marcus A. J. The role of lipids in platelet function: with particular reference to the arachidonic acid pathway. J Lipid Res. 1978 Sep;19(7):793–826. [PubMed] [Google Scholar]
  20. McGiff J. C. Cytochrome P-450 metabolism of arachidonic acid. Annu Rev Pharmacol Toxicol. 1991;31:339–369. doi: 10.1146/annurev.pa.31.040191.002011. [DOI] [PubMed] [Google Scholar]
  21. Morrison A. R., Crowley J. R. Identification of an unusual cyclooxygenase metabolite of arachidonic acid in rabbit renal medulla. Arch Biochem Biophys. 1984 Nov 1;234(2):413–417. doi: 10.1016/0003-9861(84)90287-x. [DOI] [PubMed] [Google Scholar]
  22. Morrison A. R., Pascoe N. Metabolism of arachidonate through NADPH-dependent oxygenase of renal cortex. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7375–7378. doi: 10.1073/pnas.78.12.7375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Murphy R. C., Sala A. Quantitation of sulfidopeptide leukotrienes in biological fluids by gas chromatography-mass spectrometry. Methods Enzymol. 1990;187:90–98. doi: 10.1016/0076-6879(90)87013-s. [DOI] [PubMed] [Google Scholar]
  24. Oliw E. H., Lawson J. A., Brash A. R., Oates J. A. Arachidonic acid metabolism in rabbit renal cortex. Formation of two novel dihydroxyeicosatrienoic acids. J Biol Chem. 1981 Oct 10;256(19):9924–9931. [PubMed] [Google Scholar]
  25. Powell W. S. Metabolism of prostaglandins, prostaglandin analogs and thromboxane B2 by lung and liver microsomes from pregnant rabbits. Biochim Biophys Acta. 1979 Dec 18;575(3):335–349. doi: 10.1016/0005-2760(79)90102-4. [DOI] [PubMed] [Google Scholar]
  26. Pradelles P., Grassi J., Maclouf J. Enzyme immunoassays of eicosanoids using acetylcholine esterase as label: an alternative to radioimmunoassay. Anal Chem. 1985 Jun;57(7):1170–1173. doi: 10.1021/ac00284a003. [DOI] [PubMed] [Google Scholar]
  27. Sacerdoti D., Abraham N. G., McGiff J. C., Schwartzman M. L. Renal cytochrome P-450-dependent metabolism of arachidonic acid in spontaneously hypertensive rats. Biochem Pharmacol. 1988 Feb 1;37(3):521–527. doi: 10.1016/0006-2952(88)90223-7. [DOI] [PubMed] [Google Scholar]
  28. Samuelsson B. Biosynthesis of prostaglandins. Fed Proc. 1972 Sep-Oct;31(5):1442–1450. [PubMed] [Google Scholar]
  29. Samuelsson B., Dahlén S. E., Lindgren J. A., Rouzer C. A., Serhan C. N. Leukotrienes and lipoxins: structures, biosynthesis, and biological effects. Science. 1987 Sep 4;237(4819):1171–1176. doi: 10.1126/science.2820055. [DOI] [PubMed] [Google Scholar]
  30. Scanlon M., Williams D. A., Fay F. S. A Ca2+-insensitive form of fura-2 associated with polymorphonuclear leukocytes. Assessment and accurate Ca2+ measurement. J Biol Chem. 1987 May 5;262(13):6308–6312. [PubMed] [Google Scholar]
  31. Schwartzman M. L., Abraham N. G., Carroll M. A., Levere R. D., McGiff J. C. Regulation of arachidonic acid metabolism by cytochrome P-450 in rabbit kidney. Biochem J. 1986 Aug 15;238(1):283–290. doi: 10.1042/bj2380283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schwartzman M. L., Falck J. R., Yadagiri P., Escalante B. Metabolism of 20-hydroxyeicosatetraenoic acid by cyclooxygenase. Formation and identification of novel endothelium-dependent vasoconstrictor metabolites. J Biol Chem. 1989 Jul 15;264(20):11658–11662. [PubMed] [Google Scholar]
  33. Schwartzman M., Ferreri N. R., Carroll M. A., Songu-Mize E., McGiff J. C. Renal cytochrome P450-related arachidonate metabolite inhibits (Na+ + K+)ATPase. Nature. 1985 Apr 18;314(6012):620–622. doi: 10.1038/314620a0. [DOI] [PubMed] [Google Scholar]
  34. Siegel M. I., McConnell R. T., Abrahams S. L., Porter N. A., Cuatrecasas P. Regulation of arachidonate metabolism via lipoxygenase and cyclo-oxygenase by 12-HPETE, the product of human platelet lipoxygenase. Biochem Biophys Res Commun. 1979 Aug 28;89(4):1273–1280. doi: 10.1016/0006-291x(79)92146-6. [DOI] [PubMed] [Google Scholar]
  35. Smith W. L., Marnett L. J. Prostaglandin endoperoxide synthase: structure and catalysis. Biochim Biophys Acta. 1991 Apr 24;1083(1):1–17. doi: 10.1016/0005-2760(91)90119-3. [DOI] [PubMed] [Google Scholar]
  36. Takahashi K., Capdevila J., Karara A., Falck J. R., Jacobson H. R., Badr K. F. Cytochrome P-450 arachidonate metabolites in rat kidney: characterization and hemodynamic responses. Am J Physiol. 1990 Apr;258(4 Pt 2):F781–F789. doi: 10.1152/ajprenal.1990.258.4.F781. [DOI] [PubMed] [Google Scholar]
  37. VanRollins M., Murphy R. C. Autooxidation of docosahexaenoic acid: analysis of ten isomers of hydroxydocosahexaenoate. J Lipid Res. 1984 May;25(5):507–517. [PubMed] [Google Scholar]

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