Uncovering Anti‐Inflammatory Mechanisms of <i>Elaeagnus angustifolia</i> Extract via Metabolomic and Phytochemical Analyses

Inflammation is a vital immune response that eliminates harmful stimuli. Building on our previous findings demonstrating the anti-inflammatory activity of Elaeagnus angustifolia (E. ang) aqueous crude extract at 20-80 µg/mL, this study explores its underlying mechanisms and potential biomarkers. Phytochemical profiling via liquid chromatography-tandem mass spectrometry (LC-MS/MS) identified key compounds, primarily quercetin and oleic acid. To further investigate its effects, we used an LC-MS-based metabolomics approach to analyze metabolic alterations in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells treated with 80 µg/mL of the extract. The treatment significantly altered cellular metabolism, revealing 156 differential metabolites, including amino acids, lipids, and nucleotides (pyrimidines and purines). Pathway analysis showed modulation of several metabolic routes, notably the TCA cycle, beta-oxidation, tyrosine metabolism, niacinamide salvage, and nucleotide metabolism. These results highlight the extract's capacity to modulate inflammation-related pathways at the metabolic level. Overall, our findings support the potential of E. ang aqueous extract as an antioxidant and anti-inflammatory agent. Furthermore, the study demonstrates the utility of metabolomics as a powerful tool for elucidating therapeutic mechanisms and identifying bioactive compounds in natural products.