Generation of 8-epiprostaglandin F2alpha by human monocytes. Discriminate production by reactive oxygen species and prostaglandin endoperoxide synthase-2.
F2-isoprostanes are free radical-catalyzed products of arachidonic acid. One of these compounds, 8-epiprostaglandin F2alpha (8-epi-PGF2alpha), is a mitogen and vasoconstrictor. We have shown that 8-epi-PGF2 alpha, unlike other F2-isoprostanes, is a minor product of the prostaglandin endoperoxide synthase-1 (PG G/H S-1) expressed in human platelets (Praticó, D., Lawson, J. A., and Fitzgerald, G. A. (1995) J. Biol. Chem. 270, 9800-9808). Human monocytes express PG G/H S-1 constitutively and exhibit regulated expression of PG G/H S-2. Induction of PG G/H S-2 by concanavalin A, the phorbol ester, phorbol 12-myristate 13-acetate, and bacterial lipopolysaccharide was confirmed with a specific antibody in monocytes pretreated with aspirin to inhibit PG G/H S-1. Induction of PG G/H S-2 by all three stimuli coincided with increased formation of prostaglandin E2 (PGE2), thromboxane B2 (TxB2), and 8-epi-PGF2 alpha, but not of other F2-isoprostanes. Confirmation of PG G/H S-2 as the source of 8-epi-PGF2 alpha formation was obtained by down-regulating the enzyme with dexamethasone; preventing protein synthesis with cycloheximide; and preventing synthesis of PGE2, TxB2, and 8-epi-PGF2 alpha with the specific PG G/H S-2 inhibitor, L 745,337. Monocytes also exhibit the facility to generate 8-epi-PGF2 alpha in a free radical-dependent manner. Thus, stimulation with opsonized zymosan or coincubation with low density lipoprotein was unassociated with product formation. However, coincubation of low density lipoprotein with zymosan-stimulated human monocytes resulted in marked formation of 8-epi-PGF2alpha, but not of PGE2 or TxB2. Production of 8-epi-PGF2 alpha coincided with that of thiobarbituric acid-reactive substances and lipid hydroperoxides, but was unaccompanied by PG G/H S-2 induction. Pretreatment of monocytes with the antioxidant, butylated hydroxytoluene or with superoxide dismutase, but not with L 745,337, suppressed formation of 8-epi-PGF2alpha, thiobarbituric acid-reactive substances, and lipid hydroperoxides. In conclusion, human monocytes may form bioactive 8-epi-PGF2alpha either via free radical- or enzyme-catalyzed pathways. 8-Epi-PGF2alpha is a more abundant product of monocyte PG G/H S-2 than of platelet PG G/H S-1. Formation by inducible PG G/H S-2 must be considered as a source of this compound in vivo.
Praticó
D
D
Center for Experimental Therapeutics, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
FitzGerald
G A
GA
D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D015237:Dinoprost / Q000031:analogs & derivatives* / Q000096:biosynthesis; D005938:Glucocorticoids / Q000494:pharmacology; D006801:Humans; D007189:Indans / Q000494:pharmacology; D007527:Isoenzymes / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008077:Lipoproteins, LDL / Q000378:metabolism; D009000:Monocytes / Q000378:metabolism*; D010084:Oxidation-Reduction; D011449:Prostaglandin Endoperoxides / Q000096:biosynthesis*; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism*; D011493:Protein Kinase C / Q000502:physiology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D016861:Cyclooxygenase Inhibitors; D005938:Glucocorticoids; D007189:Indans; D007527:Isoenzymes; C093700:L 745337; D008070:Lipopolysaccharides; D008077:Lipoproteins, LDL; D011449:Prostaglandin Endoperoxides; D011500:Protein Synthesis Inhibitors; D017382:Reactive Oxygen Species; C075750:8-epi-prostaglandin F2alpha; D003907:Dexamethasone; D003513:Cycloheximide; D015237:Dinoprost; D011451:Prostaglandin-Endoperoxide Synthases; D013482:Superoxide Dismutase; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate