Systematic Metabolomic Analysis of Eicosanoids after Omega-3 Polyunsaturated Fatty Acid Supplementation by a Highly Specific Liquid Chromatography–Tandem Mass Spectrometry-Based Method

Abstract

Omega-3 (ω-3) polyunsaturated fatty acids (PUFAs) have beneficial effects in many pathological processes, especially cardiovascular disease, and their protective eicosanoid metabolites are thought to play important roles. However, how ω-3 PUFAs affect the eicosanoid profile has not been elucidated comprehensively. Here, we systematically analyzed the eicosanoid metabolites induced by ω-3 PUFA supplementation. We developed an LC-MS/MS-based method covering 32 arachidonic acid (ARA) metabolites and 37 ω-3 PUFA-derived products. The limits of detection for eicosanoids were between 0.0625 and 1 pg and the detection specificity was optimized. We then quantified eicosanoids in mouse and human plasma and mouse aorta samples after ω-3 PUFA supplementation. Levels of EPA hydroxyl products, 4-HDoHE, 17,18-EEQ, 17,18-DiHETE, TXB2, and LXA4 were significantly changed in both mouse samples, and those of 2-series PGs, EDPs and DHA hydroxyl products were changed in aorta samples. Correlation network analysis of mouse plasma data revealed that some eicosanoids had higher connection degree or betweenness centrality score than others after ω-3 PUFA supplementation. Eicosanoids in human plasma were profiled across five time points after ω-3 PUFA supplementation. Fuzzy c-mean clustering algorithm suggested that the time curves of eicosanoid activity could be described with three kinetic patterns: sustained upregulation, short-term upregulation, and downregulation. This is the first systematic profiling of eicosanoids with ω-3 PUFA supplementation. The highly specific eicosanoid metabolomic and related data analysis methods would be powerful tools for comprehensive eicosanoid study.

Soluble epoxide hydrolase is involved in the development of atherosclerosis and arterial neointima formation by regulating smooth muscle cell migration

“…EET and 14,15-dihydroxyeicosatrienoic acid (DHET) were measured as previously reported (26). Briefly, cell lysate samples were spiked with 1 l of internal standard mixture (5 ng for each internal standard).…”

“…The elutions were further dried and redissolved in 30% acetonitrile. 14,15-EET and its metabolite of sEH, 14,15-DHET, were measured with ultra-highperformance liquid chromatography (UPLC; Waters, Milford, MA) and a 5500 QTRAP mass spectrometer (AB Sciex, Foster City, CA) equipped with a Turbo Ion Spray electrospray ionization source (liquid chromatography coupled with tandem mass spectrometry; LC-MS/MS) as described previously (26). The DHET-to-EET ratio was used to indicate sEH activity.…”

Quantitative proteomics reveals key proteins regulated by eicosapentaenoic acid in endothelial activation

“…Previous studies have primarily focused on a limited number of receptors and signaling pathways [5,6]. Recently, some systematic studies using metabolomics [9] and RNA sequencing [18] have been performed to explore the global perspective; however, most cellular functions are mediated by proteins, and a proteomic study of the biological mechanism of EPA has been lacking. In our study, the proteomic results were consistent with the Western blotting ( Figs.…”

“…EPA and AA share the same set of metabolic enzymes [9]. AA metabolites are often thought to exert pro-inflammatory effects that may increase the risk of atherosclerosis [9].…”

“…EPA and AA share the same set of metabolic enzymes [9]. AA metabolites are often thought to exert pro-inflammatory effects that may increase the risk of atherosclerosis [9]. EPA may displace AA from the cell membrane and partially antagonize the pro-inflammatory activity of AA metabolites [10].…”