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.

RESULTS

Network pharmacology analysis identified 15 active compounds and 69 overlapping therapeutic targets, mainly enriched in pathways related to inflammation, oxidative stress, apoptosis, and neuronal survival, including MAPK, PI3K-Akt, TNF, and NF-kB signaling pathways. In vivo, TFHL treatment significantly improved locomotor recovery, alleviated histological damage, preserved mitochondrial structure, and reduced the expression of pro-inflammatory cytokines following SCI. 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.