Specificity of eicosanoid production depends on the TLR-4-stimulated macrophage phenotype

Journal of Leukocyte Biology

Edward A. Dennis¹  Paul C. Norris¹  Christopher K. Glass  and¹  Donna Reichart¹  Darren S. Dumlao¹ 

[1] Departments of Chemistry/Biochemistry, Pharmacology, and  Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California, USA

关键词: lipidomics;    prostaglandin;    cyclooxygenase;    metabolism;    coupling;    synthase;   

DOI  :  10.1189/jlb.0311153

学科分类：生理学

来源: Federation of American Societies for Experimental Biology

【 摘 要 】

Eicosanoid metabolism differs in profile and quantity between macrophages of different tissue origin and method of elicitation, as well as between primary and immortalized macrophages after activation with inflammatory stimuli. Using a lipidomic approach, we comprehensively analyzed the eicosanoids made by murine RPMs, TGEMs, BMDM, and the macrophage-like cell line RAW after stimulation with the TLR-4-specific agonist KLA. Direct correlation among total COX metabolites, COX side-products (11-HETE, 15-HETE), COX-2 mRNA, and protein at 8 h was found when comparing each cell type. Comprehensive qPCR analysis was used to compare relative transcript levels between the terminal prostanoid synthases themselves as well as between each cell type. Levels of PGE2, PGD2, and TxB2 generally correlated with enzyme transcript expression of PGES, PGDS, and TBXS, providing evidence of comparable enzyme activities. PGIS transcript was expressed only in RPM and TGEM macrophages and at an exceptionally low level, despite high metabolite production compared with other synthases. Presence of PGIS in RPM and TGEM also lowered the production of PGE2 versus PGD2 by approximately tenfold relative to BMDM and RAW cells, which lacked this enzyme. Our results demonstrate that delayed PG production depends on the maximal level of COX-2 expression in different macrophages after TLR-4 stimulation. Also, the same enzymes in each cell largely dictate the profile of eicosanoids produced depending on the ratios of expression between them, with the exception of PGIS, which appears to have much greater synthetic capacity and competes selectively with mPGES-1.

【 授权许可】

Unknown