UHPLC-Q-Exactive Orbitrap MS-based brain-gut metabolomics implicates multi-pathway modulation by Ginseng stem-leaf saponins in chronic fatigue syndrome mice.

Ginseng stem-leaf saponins (GSLS) were evaluated as potential bioactive food ingredients by characterizing brain-gut metabolic disturbances in a chronic fatigue syndrome (CFS) mouse model and assessing the regulatory effects of GSLS using ultra-high-performance liquid chromatography-Q-Exactive Orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap MS). Male C57BL/6 mice were assigned to control, model, positive control, and GSLS groups (n = 6). A long-term forced-swimming induction protocol was used to establish fatigue-like phenotypes, and fecal and brain tissue samples were collected for untargeted metabolomics with multivariate and pathway analyses. Seventeen differential metabolites were identified in brain tissue and 17 in fecal samples in association with model induction and GSLS supplementation. The altered pathways were mainly related to lipid-derived mediators and membrane-lipid remodeling, including arachidonic acid metabolism, steroid biosynthesis, glycerophospholipid metabolism, ether lipid metabolism, and primary bile acid biosynthesis, with additional involvement of energy- and redox-relevant cofactor/vitamin metabolism, such as pantothenate and CoA biosynthesis, folate-mediated one-carbon metabolism, and lipoic acid metabolism. GSLS partly normalized fatigue-like brain and fecal metabolic abnormalities, suggesting potential links to brain-gut axis regulation involving neurotransmitter-related metabolism, hypothalamic-pituitary-adrenal (HPA)-axis-associated steroid metabolism, inflammation, and oxidative stress. These findings support GSLS as a promising bioactive ingredient for maintaining vitality and metabolic resilience in functional food applications.