Cerebral ischemia resulting from middle cerebral artery occlusion (MCAO) significantly impairs neuronal function and exacerbates cognitive deficits. This study investigates the neuroprotective effects of Biochanin A, a natural isoflavonoid, in a MCAO model using Wistar rats. Fifty rats were divided into five groups: a control group, a MCAO group, and three treatment groups receiving 200 mg/kg and 400 mg/kg of Biochanin A. Cognitive function was assessed using the Morris water maze test, while serum levels of neurotrophic factors (GDNF and BDNF) and inflammatory cytokines (TNF-α, IL-1β, IL-6, IL-10, and IL-4) were measured through enzyme-linked immunosorbent assay (ELISA). Additionally, gene expression of JAK2 and STAT3 was analyzed via real-time PCR. Results indicated that MCAO significantly impaired cognitive function and increased pro-inflammatory cytokines while decreasing neurotrophic factors (p<0.05). In contrast, Biochanin A treatment improved escape latency in the Morris water maze and elevated serum GDNF and BDNF levels (p<0.05). Furthermore, Biochanin A administration significantly modulated the expression of JAK2 and STAT3, promoting neuroprotection through the JAK2/STAT3 signaling pathway. These findings suggest that Biochanin A mitigates cognitive deficits and neuroinflammation following MCAO, potentially by enhancing neurotrophic factor levels and modulating inflammatory responses. Thus, Biochanin A represents a promising therapeutic candidate for ischemic stroke recovery.

KEY WORDS: Biochanin A; Cerebral infraction; Network pharmacology; Cerebral ischemia-reperfusion.

CHARILATU, W. & JIE, X. Network pharmacology-driven elucidation of biochanin A's neuroprotective mechanism in cerebral ischemia: JAK2/STAT3 signaling pathway modulation via p-JAK2, JAK2, p-STAT3, and STAT3 regulation. Int. J. Morphol., 44(4):1263-1278, 2026.