Abstract:

Using network pharmacology combined with cellular experiments， this study investigated the neuroprotective effects of wogonin （WOG） on cerebral ischemia-reperfusion injury （CIRI） and its underlying molecular mechanisms. Potential targets of wogonin were predicted using SwissTargetPrediction， and CIRI-related genes were collected from GeneCards databases. Common targets were identified for GO and KEGG enrichment analyses and for constructing a protein-protein interaction network. Molecular docking was performed to evaluate the binding affinity between wogonin and key targets. PC12 cells were used for in vitro experiments and divided into a control group， an oxygen-glucose deprivation （OGD） group， and an OGD+WOG group. Colony formation assays were conducted to assess cell proliferation. FerroOrange and C11 BODIPY 581/591 probes were used to detect Fe2⁺ levels and lipid peroxidation， respectively. Immunofluorescence was used to measure the levels of glutathione peroxidase 4 （GPX4） and solute carrier family 7 member 11 （SLC7A11）. A GSH/GSSG assay kit was used to quantify glutathione （GSH） and Western blotting was performed to evaluate arachidonate 15-lipoxygenase （ALOX15） protein expression. The results showed that a total of 51 common targets of WOG and CIRI were identified. GO and KEGG analyses revealed significant enrichment in arachidonic acid metabolism. Molecular docking showed strong binding between WOG and ALOX 15 （binding energy -8.6 kcal·mol^-1）. Cell experiments demonstrated that WOG significantly alleviated OGD-induced suppression of colony formation， reduced Fe2⁺ accumulation and lipid peroxidation， increased GSH levels， upregulated GPX4 and SLC7A11， and downregulated ALOX15. In conclusion， wogonin may alleviate CIRI by inhibiting OGD-induced ferroptosis in PC12 cells via targeting ALOX15.

Key words: ferroptosis, wogonin, cerebral ischemia-reperfusion injury, network pharmacology, molecular docking, ALOX15