Pro-resolving actions and stereoselective biosynthesis of 18S E-series resolvins in human leukocytes and murine inflammation
Sungwhan F. Oh, … , Rong Yang, Charles N. Serhan
Sungwhan F. Oh, … , Rong Yang, Charles N. Serhan
Published January 4, 2011
Citation Information: J Clin Invest. 2011;121(2):569-581. https://doi.org/10.1172/JCI42545.
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Research Article Immunology

Pro-resolving actions and stereoselective biosynthesis of 18S E-series resolvins in human leukocytes and murine inflammation

Abstract

E-series resolvins are antiinflammatory and pro-resolving lipid mediators derived from the ω-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA) that actively clear inflammation to promote tissue homeostasis. Aspirin, in addition to exerting antithrombotic actions, also triggers the biosynthesis of these specialized pro-resolving mediators. Here, we used metabolomic profiling to investigate the biosynthesis of E-series resolvins with specific chiral chemistry in serum from human subjects and present evidence for new 18S series resolvins. Aspirin increased endogenous formation of 18S-hydroxyeicosapentaenoate (18S-HEPE) compared with 18R-HEPE, a known resolvin precursor. Human recombinant 5-lipoxygenase used both enantiomers as substrates, and recombinant LTA4 hydrolase (LTA4H) converted chiral 5S(6)-epoxide–containing intermediates to resolvin E1 and 18S-resolvin E1 (RvE1 and 18S-RvE1, respectively). 18S-RvE1 bound to the leukocyte GPCRs ChemR23 and BLT1 with increased affinity and potency compared with the R-epimer, but was more rapidly inactivated than RvE1 by dehydrogenase. Like RvE1, 18S-RvE1 enhanced macrophage phagocytosis of zymosan, E. coli, and apoptotic neutrophils and reduced both neutrophil infiltration and proinflammatory cytokines in murine peritonitis. These results demonstrate two parallel stereospecific pathways in the biosynthesis of E-series resolvins, 18R- and 18S-, which are antiinflammatory, pro-resolving, and non-phlogistic and may contribute to the beneficial actions of aspirin and ω-3 polyunsaturated fatty acids.

Authors

Sungwhan F. Oh, Padmini S. Pillai, Antonio Recchiuti, Rong Yang, Charles N. Serhan

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Figure 6

Molecular pathways of 18S-RvE1 actions: ligand-receptor interaction and metabolic inactivation.

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Molecular pathways of 18S-RvE1 actions: ligand-receptor interaction and ...
(A) ChemR23 activation. Increasing doses of RvE1 (diamonds) or 18S-RvE1 (squares) were incubated with the ChemR23-overexpressing reporter system, followed by β-gal substrate incubation. Luminescence was measured to obtain the RvE1 EC50, 1.37 × 10–10 M, and 18S-RvE1 EC50, 6.33 × 10–12 M, from fitted dose-response curves. (B) BLT1 antagonism. BLT1-overexpressing cells were incubated with different concentrations of 18S-RvE1 or RvE1, followed by 30 nM LTB4, and further incubated for 30 minutes. Results are mean ± SEM; n ≥ 3. P < 0.05, RvE1 versus 18S-RvE1. (C) Comparison between RvE1 and 18S-RvE1 by 15-PGDH–mediated oxidation (diamonds: RvE1, squares: 18S-RvE1) addressed in the presence of NAD+ as a cofactor. NADH was monitored as a readout of dehydrogenation. Results shown are representative of n = 3 with similar trends.