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

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.