Abstract
Metabolism of arachidonic acid results in a host of biologically active compounds with profound effects on airway inflammation (1). After activation of cellular phospholipases and release of free arachidonic acid, catalyzed insertion of oxygen occurs enzymatically via action of one of the two known cyclooxygenase isoenzymes (COX-1 and COX-2). The unstable bicyclic intermediate, PGH2, undergoes subsequent metabolism to form prostaglandins (PG), thromboxane (Tx), and leukotrienes (LT) (see Fig.1). In addition, free radicals can oxygenate arachidonate although it is bound to the diacylgycerol backbone of membrane phospholipids. The family of compounds formed in this way, known as isoprostanes, are stereochemically different and incorporate a large number of regioisomeric compounds that may confound measurement of PG (2–5 and see Chapter 33). Arachidonic acid can also be metabolized by specific cytochrome P450 enzymes to regioisomeric epoxides and stereo specific hydro xyeicosatetraenoic (HETE) acids (6).
Overview of pathways of metabolism of arachidonic acid during airway inflammation. Following activation of cellular phospholipases, arachidonic acid is cleaved from membrane phospholipids. It is undergoing dioxygenation catalyzed by cyclooxygenase (either COX-1 or COX-2 isoforms) to form the unstable endoperoxide intermediate PGH2. Specific isomerases with varied cellular distribution further metabolize PGH2 to bioactive prostaglandins and thromboxanes
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References
Peters-Golden, M. (1997) Lipid mediator syntheis by lung macrophages, in Lung Macrophages and Dendritic Cells in Health and Disease (Lipscomb, M. F. and Russell, S. W., ed.) 102, 151–182.
Roberts, L. J., 2nd., Brame, C. J., Chen, Y., and Morrow, J. D. (1999) Novel eicosanoids. Isoprostanes and related compounds. Methods in Mol. Bio. 120, 257–285.
Roberts, L. J., 2nd., Salomon, R. G., Morrow, J. D., and Brame, C. J. (1999) New developments in the isoprostane pathway: identification of novel highly reactive gamma-ketoaldehydes (isolevuglandins) and characterization of their protein adducts. FASEB Journal. 13, 1157–1168.
Morrow, J. D. and Roberts, L. J. 2nd. (1999) Mass spectrometric quantification of F2-isoprostanes in biological fluids and tissues as measure of oxidant stress. Methods in Enzymology. 300, 3–12.
Dworski, R., Murray, J. J, Roberts, L. J., II, Oates, J. A., Morrow, J. D., Fisher, L., and Sheller, J. R. (1999) Allergen-induced Synthesis of F2-Isoprostanes in Atopic Asthmatics Evidence for Oxidant Stress Am. J. Respir. Crit. Care Med. 160, 1947–1951.
Capdevila, J., Falck, J., and Estabrook, R. (1992) Cytochrome P450 and the arachidonate cascade. FASEB J 6, 731–736.
Murray, J. J., Tonnel, A. B., Brash, A. R., Roberts, L. J., 2nd, and Gosset, P. (1986) Workman R. Capron A. Oates JA. Release of prostaglandin D2 into human airways during acute antigen challenge. N. Engl. J. of Med. 315, 800–804.
Luppinetti, M. D., Sheller, J. R., Catella, F., FitzGerald, G. A. (1989) Thrombox-ane biosynthesis in allergen induced bronchospasm: evidence for platelet activation. Am. Rev. Respir. Dis. 140, 932–935.
Christman, B. W., Gay, J. C., Christman, J. W., Prakash, C., Blair, I. A. (1991) Analysis of effector cell-derived lyso platelet activating factor by electron capture negative ion mass spectrometry. Biol. Mass Spectrom. 20, 545–552.
Christman, B. W., Christman, J. W., Dworski, R., Blair, I. A., Prakash, C. (1993) PGE2 limits arachidonic acid availability and inhibits LTB4 synthesis in alveolar macrophages by a nonphospholipase A2 mechanism. J. Immunol. 151, 2096–2104.
Sebaldt, R., Sheller, J., Oates, J., Roberts, L. J., II, and Fitzgerald, G. (1990) Inhibition of eicosanoid biosynthesis by glucocorticoids in humans. Proc. Natl. Acad. Sci. USA 87, 6974–6978.
Morrow, J. D., Prakash, C., Awad, J. A., Duckworth, T. A., Zackert, W. E., Blair, I. A., Oates, J. A., and Roberts, L. J., II. (1991) Quantification of the major urinary metabolite of prostaglandin D2 by a stable isotope dilution mass spectrometric assay. Analyt. Biochem. 193, 142–148.
Daniel, V. C., Minton, T. A., Brown, N. J., Nadequ, J. H., and Morrow, J. D. (1994) Simplified assay for the quantification of 2,3-dinor-6-keto-prostaglandin F1a by gas chromatography-mass spectrometry. J. Chromatography B: Biomedi-cal Applications 653(2), 117–122.
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Dworski, R., Sheller, J.R., Christman, B.W. (2001). Quantitative Analysis of Cyclooxygenase Metabolites of Arachidonic Acid. In: Rogers, D.F., Donnelly, L.E. (eds) Human Airway Inflammation. Methods in Molecular Medicine, vol 56. Humana Press. https://doi.org/10.1385/1-59259-151-5:411
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DOI: https://doi.org/10.1385/1-59259-151-5:411
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