Contributions of the Three CYP1 Monooxygenases to Pro-Inflammatory and Inflammation-Resolution Lipid Mediator Pathways

• Published in: The Journal of immunology (1950) Vol. 191; no. 6; pp. 3347 - 3357
• Main Authors: Divanovic, Senad, Dalli, Jesmond, Jorge-Nebert, Lucia F, Flick, Leah M, Gálvez-Peralta, Marina, Boespflug, Nicholas D, Stankiewicz, Traci E, Fitzgerald, Jonathan M, Somarathna, Maheshika, Karp, Christopher L, Serhan, Charles N, Nebert, Daniel W
• Format: Journal Article
• Language: English
• Published: United States 15.09.2013
• Subjects: Animals; Cytochrome P-450 Enzyme System - immunology; Cytochrome P-450 Enzyme System - metabolism; Humans; Inflammation - immunology; Inflammation - metabolism; Inflammation Mediators - immunology; Inflammation Mediators - metabolism; Lipids - immunology; Metabolome; Mice; Mice, Inbred C57BL; Mice, Knockout; Neutrophils - immunology; Neutrophils - metabolism; Signal Transduction - immunology
• ISSN: 0022-1767; 1550-6606; 1550-6606
• DOI: 10.4049/jimmunol.1300699

All three cytochrome P450 1 (CYP1) monooxygenases are believed to participate in lipid mediator biosynthesis and/or their local inactivation; however, distinct metabolic steps are unknown. We used multiple-reaction monitoring and liquid chromatography-UV coupled with tandem mass spectrometry–based lipid-mediator metabololipidomics to identify and quantify three lipid-mediator metabolomes in basal peritoneal and zymosan-stimulated inflammatory exudates, comparing Cyp1a1/1a2/1b1(−/−) C57BL/6J-background triple-knockout mice with C57BL/6J wild-type mice. Significant differences between untreated triple-knockout and wild-type mice were not found for peritoneal cell number or type or for basal CYP1 activities involving 11 identified metabolic steps. Following zymosan-initiated inflammation, 18 lipid mediators were identified, including members of the eicosanoids and specialized proresolving mediators (i.e., resolvins and protectins). Compared with wild-type mice, Cyp1 triple-knockout mice exhibited increased neutrophil recruitment in zymosan-treated peritoneal exudates. Zymosan stimulation was associated with eight statistically significantly altered metabolic steps: increased arachidonic acid–derived leukotriene B4 (LTB4) and decreased 5S-hydroxyeicosatetraenoic acid; decreased docosahexaenoic acid–derived neuroprotectin D1/protectin D1, 17S-hydroxydocosahexaenoic acid, and 14S-hydroxydocosahexaenoic acid; and decreased eicosapentaenoic acid–derived 18R-hydroxyeicosapentaenoic acid (HEPE), 15S-HEPE, and 12S-HEPE. In neutrophils analyzed ex vivo, elevated LTB4 levels were shown to parallel increased neutrophil numbers, and 20-hydroxy–LTB4 formation was found to be deficient in Cyp1 triple-knockout mice. Together, these results demonstrate novel contributions of CYP1 enzymes to the local metabolite profile of lipid mediators that regulate neutrophilic inflammation.