Molecular mechanism of total flavonoids of hawthorn leaves in spinal cord injury treatment: A combined network pharmacology and experimental study.

BACKGROUND AND OBJECTIVE: Spinal cord injury (SCI) is a devastating neurological disorder characterized by irreversible motor and sensory dysfunction, largely driven by secondary injury mechanisms including oxidative stress, inflammation, and apoptosis. Total flavonoids of hawthorn leaves (TFHL), a traditional Chinese medicine, have demonstrated antioxidant and anti-inflammatory properties; however, their therapeutic effects and mechanisms in SCI remain unclear. METHODS: Network pharmacology was employed to identify the active compounds, potential targets, and key signaling pathways of TFHL in SCI through target prediction, protein-protein interaction analysis, and GO and KEGG enrichment analyses. Experimental validation was performed using a rat SCI model, LPS-stimulated BV-2 microglial cells, and RAW264.7 macrophages. Functional recovery was assessed by BBB scoring, while histopathological changes, ultrastructural alterations, inflammatory cytokine expression, and MAPK pathway activation were evaluated using H&E staining, transmission electron microscopy, RT-qPCR, and western blotting. In vitro, TFHL markedly suppressed LPS-induced activation of the MAPK pathway in microglial cells. CONCLUSION: TFHL improves functional and histopathological outcomes in SCI models and attenuates inflammatory activation. These effects are consistent with multi-pathway regulation predicted by network pharmacology and are supported, at least in part, by MAPK pathway suppression, warranting further mechanistic and translational studies.