Integrated Approach Reveals Fermented Moringa oleifera Leaves Extracts’ Impact on Mouse Sleep

Sleep disturbances are linked to metabolic and neurological dysregulation. <i>Moringa oleifera</i> leaves, rich in bioactive compounds, may improve sleep via gut-brain axis modulation. This study investigated the sleep-enhancing effects of fermented <i>Moringa oleifera</i> leaf extract (FM) in mice using metabolomics, gut microbiota analysis, network pharmacology, and molecular docking. A 1:1 combination of Lactobacillus plantarum GDMCC 1.2685 and <i>L. swissii</i> GDMCC 1.791 optimally fermented FM, increasing GABA by 1.67-fold and total amino acids to 46,058.20 ± 845.53 μg/g. FM shortened sleep latency, increased sleep duration, and elevated brain GABA while reducing glutamate (Glu) and Glu/GABA ratios. Hypothalamic metabolomics identified seven sleep-related metabolites, implicating glycerophospholipid, tryptophan, and purine metabolism pathways. FM also reduced <i>Mycobacterium anisopliae</i> (a gut bacterium associated with insomnia) and increased the Firmicutes/Bacteroidetes ratio. Network pharmacology revealed that FM's effects were mediated via GABA, Glu, and serotonin (5-HT) pathways. These findings demonstrate that FM improves sleep by modulating hypothalamic neurotransmitters and gut microbiota, exerting sedative-hypnotic effects through amino acid, purine, and energy metabolism.