To investigate the neuroprotective effect of vccarin on ischemic brain injury in mice.

Potential targets of vaccarin and ischemic brain injury were predicted using databases such as GeneCard, Swiss Target Prediction, Targetnet, Pharmmapper, and TTD (search date: October 30, 2024). Intersecting targets were identified and analyzed via protein-protein interaction (PPI) network construction (STRING database; visualization with Cytoscape). Functional enrichment analysis (GO and KEGG pathways) was performed using DAVID and MicrobeDB. Molecular docking (AutoDock Vina) assessed binding affinities between vaccarin and core targets (degree ≥40). In animal experiments, focal cerebral ischemia was induced in mice via distal middle cerebral artery occlusion (dMCAO). Vaccarin (10, 20, 40 mg/kg) or vehicle was administered intraperitoneally for 5 days post-surgery. Neurological function scores and rotarod tests were performed on days 1, 3, and 5 post-surgery. After drug administration on day 5, triphenyltetrazolium chloride (TTC) staining and histopathological section staining were performed to verify the effects of vaccarin on ischemic brain injury in mice.

After removing duplicates and obtaining the intersection of the targets, a total of 63 overlapping targets of vaccarin and ischemic stroke (IS) were identified. The PPI network of the intersecting targets was constructed using the Sting platform. GO and KEGG enrichment analysis indicated that vaccarin primarily regulated biological processes such as hypoxia response, inflammation, and apoptosis, and Highlighted VEGF and IL-17 signaling pathways. Molecular docking results showed that the binding energies of vaccarin with core targets (degree≥40) were strong, with binding energies all less than -7 kcal/mol. In animal experiments, vaccarin significantly reduced the mNSS neurological scores and prolonged the time spent on the rotarod in ischemic mice, with statistically significant differences (P<0.05 or P<0.01). Compared to the sham group, vaccarin intervention significantly reduced the infarct volume in mice, with statistical significance (P<0.05 or P<0.01). Pathological staining showed that vaccarin significantly alleviated the loss of neurons and improved neuronal morphology. The loss of Nissl bodies was also notably reduced.

Vaccarin confers neuroprotection in ischemic brain injury by modulating hypoxia-inflammatory-apoptosis pathways via multi-target interactions, with VEGF and IL-17 signaling as key mechanistic components.