Identification and signature profiles for pro-resolving and inflammatory lipid mediators in human tissue

Abstract

Resolution of acute inflammation is an active process locally controlled by a novel genus of specialized pro-resolving mediators (SPM) that orchestrate key resolution responses. Hence, it is of general interest to identify individual bioactive mediators and profile their biosynthetic pathways with related isomers as well as their relation(s) to classic eicosanoids in mammalian tissues. Lipid mediator (LM)-SPM levels and signature profiles of their biosynthetic pathways were investigated using liquid chromatography-tandem mass spectrometry (LC-MS-MS)-based LM metabololipidomics. LM and SPM were identified using ≥6 diagnostic ions and chromatographic behavior matching with both authentic and synthetic materials. This approach was validated using the composite reference plasma (SRM1950) of 100 healthy individuals. Using targeted LM metabololipidomics, we profiled LM and SPM pathways in human peripheral blood (plasma and serum) and lymphoid organs. In these, we identified endogenous SPM metabolomes, namely, the potent lipoxins (LX), resolvins (Rv), protectins (PD), and maresins (MaR). These included RvD1, RvD2, RvD3, MaR1, and NPD1/PD1, which were identified in amounts within their bioactive ranges. In plasma and serum, principal component analysis (PCA) identified signature profiles of eicosanoids and SPM clusters. Plasma-SPM increased with omega-3 and acetylsalicylic acid intake that correlated with increased phagocytosis of Escherichia coli in whole blood. These findings demonstrate an approach for identification of SPM pathways (e.g., resolvins, protectins, and maresins) in human blood and lymphoid tissues that were in amounts commensurate with their pro-resolving, organ protective, and tissue regeneration functions. LM metabololipidomics coupled with calibration tissues and physiological changes documented herein provide a tool for functional phenotypic profiling.

Protectin D1 n-3 DPA and resolvin D5 n-3 DPA are effectors of intestinal protection

“…Of note, levels of these mediators were within their bioactive concentration (femtomolar to nanomolar) (13,14), a feature aligned with recent findings where SPM at bioactive concentrations were identified in human tissues including spleen, lymph nodes, plasma (25), and also milk (26). The protective role of endogenous SPM was tested using a 15-lipoxygenase inhibitor that decreased the formation of PD1 n-3 DPA and that was associated with an increase in the production of proinflammatory molecules and colon inflammation.…”

“…Human and mouse gastrointestinal tissues extraction and LM metabololipidomics was conducted as described previously (25).…”

“…Intestinal levels of cytokines were assayed using ELISA kit. Colonic samples were harvested for metabololipidomics analysis, as previously described (25).…”

Effect of Eicosapentaenoic and Docosahexaenoic Acids Added to Statin Therapy on Coronary Artery Plaque in Patients With Coronary Artery Disease: A Randomized Clinical Trial

“…EPA and DHA supplementation for 6 months in post–myocardial infarction patients was associated with a favorable effect on left ventricular remodeling and a significant reduction in noninfarct myocardial fibrosis measured by cardiac magnetic resonance imaging (mean reduction, −5.6%; 95% CI, −10.4% to −0.9%; P =0.022) compared with placebo 35. In a subset of 6 patients from the current study, several resolvins, which have been shown to be present in healthy subjects,36, 37, 38 were absent or present at low levels in CAD patients not on omega‐3 ethyl‐ester; the resolvins were restored or their level increased in those receiving omega‐3 ethyl‐ester 39. The stimulation of resolution of infection and inflammation by resolvins (the downstream products of EPA and DHA) may be responsible for the significantly lower rate of nonserious infectious disease events in the omega‐3 group 40, 41.…”

Resolvin D1 Reduces Emphysema and Chronic Inflammation

“…Left lungs were each placed in cold methanol and extracted as described followed by targeted lipid mediatoreSPM profiling using liquid chromatography with tandem mass spectrometry. 14,15 Western Blot Analysis Sigma-Aldrich). Protein concentration was quantitated using a bicinchoninic acid detection assay (Pierce, Rockford, IL).…”