Screening of fish oil fatty acids for antihyperlipidemic activity based on network pharmacology and validation of synergistic efficacy in vitro

Fish oil, rich in docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), shows promise as an anti-hyperlipidemic agent. We explored the synergistic lipid-lowering effects between long-chain monounsaturated fatty acids, EPA, and DHA using network pharmacology and experimental validation in HepG2 cells. Various fish oils, particularly Engraulis japonicus and Pacific saury, effectively reduced oleic acid-induced lipid accumulation. Network pharmacology identified 64 potential targets for fish oil fatty acids' synergistic anti-hyperlipidemic action. Validation revealed their superior impact on upregulating CPT1A and RXRA mRNA, promoting lipid metabolism, and suppressing SCD and FABP1 mRNA, inhibiting lipid synthesis. This combination also reduced IL6 expression, maintaining lipid homeostasis. Activating AMPK and PPAR pathways, it achieved lipid-lowering effects. These findings highlight the potential of fish oil fatty acids for anti-hyperlipidemic interventions, offering a novel research perspective.