Characterization and identification of cytochrome P450 metabolites of arachidonic acid released by human peritoneal macrophages obtained from the pouch of Douglas

• Published in: Prostaglandins, leukotrienes and essential fatty acids leukotrienes and essential fatty acids, 2002-12, Vol.67 (6), p.397-404
• Main Authors: Werner, K., Schaefer, W.R., Schweer, H., Deppert, W.R., Karck, U., Zahradnik, H.P.
• Format: Article
• Language: English
• Subjects: Arachidonic Acid - analysis; Arachidonic Acid - chemistry; Arachidonic Acid - metabolism; Biological and medical sciences; Chromatography, High Pressure Liquid; Cytochrome P-450 Enzyme System - metabolism; Douglas' Pouch - pathology; Female; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Gas Chromatography-Mass Spectrometry; Humans; Hydroxyeicosatetraenoic Acids - analysis; Hydroxyeicosatetraenoic Acids - chemistry; Hydroxyeicosatetraenoic Acids - metabolism; Immunobiology; Isotope Labeling; Macrophages, Peritoneal - enzymology; Macrophages, Peritoneal - metabolism; Molecular Conformation; Monocytes, macrophages; Myeloid cells: ontogeny, maturation, markers, receptors
• ISSN: 0952-3278; 1532-2823
• DOI: 10.1054/plef.2002.0449

Cytochrome P450 metabolism of arachidonic acid (AA) was investigated in human peritoneal macrophages which play a central role in chronic pelvic diseases in women (for example in endometriosis). The formation of eicosanoids other than prostaglandins (PGs) by these cells is still unknown. In non-activated macrophages obtained from women in the reproductive age, the main [ 3H]-AA metabolites coeluted with epoxyeicosatrienoic acids, dihydroxyeicosatrienoic acids (DHETs) and hydroxyeicosatetraenoic acids (HETEs) in reverse-phase HPLC. After zymosan activation a shift to PGs pathway was observed. Treatment with low doses of 2,3,7,8-tetrachlorodibenzo- p -dioxin increased the formation of a metabolite coeluting with 5,6-DHET. By gas chromatography/mass spectrometry 5,6-DHET (after β-naphthoflavone induction), and 14,15-DHET as well as 11,12-DHET (after AA stimulation) were identified as major epoxygenase metabolites, respectively. The enantioselective formation of 12( S)-HETE was demonstrated by chiral-phase HPLC. Our findings demonstrate that non-activated peritoneal macrophages produce substantial amounts of bioactive cytochrome P450 metabolites of AA.