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. 1982 Jun 1;155(6):1653–1664. doi: 10.1084/jem.155.6.1653

Uptake and metabolism of monohydroxy-eicosatetraenoic acids by macrophages

PMCID: PMC2186712  PMID: 6951919

Abstract

Within 5 min, resting macrophages metabolize microM quantities of exogenous arachidonic acid (20:4) to cyclooxygenase and lipoxygenase products. Mono-HETEs represent a major class of metabolites recovered from the medium. However, the quantity of mono-Hetes progressively decreases over a 60-min incubation period, with a concomitant increase in more polar lipoxygenase products, suggesting additional metabolic fates for these hydroxy acids. This was directly confirmed by exposing resident macrophage cultures to radiolabeled 15-, 12-, and 5-HETEs (1 microM). 12-30% of the recovered HETEs were cell-associated and predominantly esterified into phospholipid. High pressure liquid chromatography analyses of medium extracts indicated that 50% of each HETE was also converted to 10 or more metabolites over a 60-min time- course, a rate slower than for 20:4. The major metabolite generated from each mono-HETE had the elution characteristics of a di-HETE. The 5- HETE product has a triene spectrum similar to that of 5(S), 12(S)-di- HETE, whereas the 15- and 12-HETE products exhibited single ultraviolet absorption maxima, indicating a metabolic pathway for 5-HETE distinct from the other mono-HETEs. None of the stable cyclooxygenase products of 20:4 (6-keto PGF1 alpha, PGF2 alpha, PGE2, TXB2) nor polar metabolites of mono-HETEs are either incorporated or metabolized. The results indicate that macrophages have the capacity to specifically metabolize 20:4 and mono-HETEs to polar oxygenated products in the absence of a discernible trigger.

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Selected References

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  1. Alam I., Ohuchi K., Levine L. Determination of cyclooxygenase products and prostaglandin metabolites using high-pressure liquid chromatography and radioimmunoassay. Anal Biochem. 1979 Mar;93(2):339–345. doi: 10.1016/s0003-2697(79)80160-8. [DOI] [PubMed] [Google Scholar]
  2. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  3. Bokoch G. M., Reed P. W. Stimulation of arachidonic acid metabolism in the polymorphonuclear leukocyte by an N-formylated peptide. Comparison with ionophore A23187. J Biol Chem. 1980 Nov 10;255(21):10223–10226. [PubMed] [Google Scholar]
  4. Borgeat P., Samuelsson B. Arachidonic acid metabolism in polymorphonuclear leukocytes: effects of ionophore A23187. Proc Natl Acad Sci U S A. 1979 May;76(5):2148–2152. doi: 10.1073/pnas.76.5.2148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Borgeat P., Samuelsson B. Metabolism of arachidonic acid in polymorphonuclear leukocytes. Structural analysis of novel hydroxylated compounds. J Biol Chem. 1979 Aug 25;254(16):7865–7869. [PubMed] [Google Scholar]
  6. Dahlén S. E., Björk J., Hedqvist P., Arfors K. E., Hammarström S., Lindgren J. A., Samuelsson B. Leukotrienes promote plasma leakage and leukocyte adhesion in postcapillary venules: in vivo effects with relevance to the acute inflammatory response. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3887–3891. doi: 10.1073/pnas.78.6.3887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Funk M. O., Isacc R., Porter N. A. Preparation and purification of lipid hydroperoxides from arachidonic and gamma-linolenic acids. Lipids. 1976 Feb;11(2):113–117. doi: 10.1007/BF02532660. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Hansson G., Lindgren J. A., Dahlén S. E., Hedqvist P., Samuelsson B. Identification and biological activity of novel omega-oxidized metabolites of leukotriene B4 from human leukocytes. FEBS Lett. 1981 Jul 20;130(1):107–112. doi: 10.1016/0014-5793(81)80676-x. [DOI] [PubMed] [Google Scholar]
  10. Lewis R. A., Goetzl E. J., Drazen J. M., Soter N. A., Austen K. F., Corey E. J. Functional characterization of synthetic leukotriene B and its stereochemical isomers. J Exp Med. 1981 Oct 1;154(4):1243–1248. doi: 10.1084/jem.154.4.1243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lindgren J. A., Hansson G., Samuelsson B. Formation of novel hydroxylated eicosatetraenoic acids in preparations of human polymorphonuclear leukocytes. FEBS Lett. 1981 Jun 15;128(2):329–335. doi: 10.1016/0014-5793(81)80110-x. [DOI] [PubMed] [Google Scholar]
  12. Maas R. L., Brash A. R., Oates J. A. A second pathway of leukotriene biosynthesis in porcine leukocytes. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5523–5527. doi: 10.1073/pnas.78.9.5523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mahoney E. M., Hamill A. L., Scott W. A., Cohn Z. A. Response of endocytosis to altered fatty acyl composition of macrophage phospholipids. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4895–4899. doi: 10.1073/pnas.74.11.4895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rigaud M., Durand J., Breton J. C. Transfomration of arachidonic acid into 12-hydroxy-5,8,10,14-eicosatetraenoic acid by mouse peritoneal macrophages. Biochim Biophys Acta. 1979 May 25;573(2):408–412. doi: 10.1016/0005-2760(79)90074-2. [DOI] [PubMed] [Google Scholar]
  15. Rouzer C. A., Scott W. A., Cohn Z. A., Blackburn P., Manning J. M. Mouse peritoneal macrophages release leukotriene C in response to a phagocytic stimulus. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4928–4932. doi: 10.1073/pnas.77.8.4928. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rouzer C. A., Scott W. A., Hamill A. L., Cohn Z. A. Dynamics of leukotriene C production by macrophages. J Exp Med. 1980 Nov 1;152(5):1236–1247. doi: 10.1084/jem.152.5.1236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Scott W. A., Pawlowski N. A., Andreach M., Cohn Z. A. Resting macrophages produce distinct metabolites from exogenous arachidonic acid. J Exp Med. 1982 Feb 1;155(2):535–547. doi: 10.1084/jem.155.2.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Scott W. A., Pawlowski N. A., Murray H. W., Andreach M., Zrike J., Cohn Z. A. Regulation of arachidonic acid metabolism by macrophage activation. J Exp Med. 1982 Apr 1;155(4):1148–1160. doi: 10.1084/jem.155.4.1148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Scott W. A., Zrike J. M., Hamill A. L., Kempe J., Cohn Z. A. Regulation of arachidonic acid metabolites in macrophages. J Exp Med. 1980 Aug 1;152(2):324–335. doi: 10.1084/jem.152.2.324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Stenson W. F., Parker C. W. 12-L-hydroxy-5,8,10,14-eicosatetraenoic acid, a chemotactic fatty acid, is incorporated into neutrophil phospholipids and triglyceride. Prostaglandins. 1979 Aug;18(2):285–292. doi: 10.1016/0090-6980(79)90115-1. [DOI] [PubMed] [Google Scholar]
  21. Stenson W. F., Parker C. W. Metabolism of arachidonic acid in ionophore-stimulated neutrophils. Esterification of a hydroxylated metabolite into phospholipids. J Clin Invest. 1979 Nov;64(5):1457–1465. doi: 10.1172/JCI109604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Unger W. G., Stamford I. F., Bennett A. Extraction of prostaglandins from human blood. Nature. 1971 Oct 1;233(5318):336–337. doi: 10.1038/233336b0. [DOI] [PubMed] [Google Scholar]

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