Integrating network pharmacology, molecular docking and experimental verification to explore the protective effect of Buyang Huanwu Decoction on cerebral ischemic stroke

Cerebral ischemic stroke (CIS), the most prevalent form of stroke worldwide, exerts substantial impacts on both public health and socioeconomic conditions, particularly among elderly populations. Buyang Huanwu Decoction (BYHWD) has been recognized as a representative herbal formula for the management of ischemic stroke; however, its underlying cellular and molecular mechanisms remain to be fully elucidated. Through network pharmacology prediction and in vitro experimental validation, this study elucidated the BYHWD and its active constituent, quercetin, inhibit the oxygen-glucose deprivation/reperfusion (OGD/R) injury model in HT22 neuronal cells, thereby providing experimental evidence supporting its therapeutic application in CIS. Network pharmacology analysis revealed that BYHWD constituents, including quercetin, kaempferol, stigmasterol, β-sitosterol, and baicalein, exerted their therapeutic effects through multiple signaling pathways such as the PI3K-Akt, IL-17, and TNF signaling pathways, interacting with key protein targets including AKT1, IL6, TNF-α, IL1β, and TP53. The CCK-8 assay results demonstrated that treatment with BYHWD extract and quercetin significantly improved the OGD/R-induced HT22 cells reductions in cell viability ( p  < 0.05), with effects showing a concentration-dependent trend observed within a specific concentration range. Further investigations confirmed a direct interaction between quercetin and TNF-α via biomolecular interaction analysis. Moreover, both high, medium, and low concentrations of BYHWD and quercetin significantly reduced TNF-α protein expression in the OGD/R-induced HT22 cells ( p  < 0.05). Western blot analysis corroborated the inhibitory effect of quercetin on TNF-α protein expression under the same experimental conditions. BYHWD demonstrates a multi-component, multi-target, and multi-pathway mechanism of action in the treatment of CIS. TNF-α and quercetin are identified as key therapeutic targets and active components, respectively. Both BYHWD and quercetin effectively mitigate TNF-α-mediated inflammatory responses and ameliorate cellular injury in the OGD/R-induced HT22 cells.