Mechanistic investigation of Xijiao Dihuang decoction for ischemic stroke: A network pharmacology and in vivo validation study

Ischemic stroke is the most prevalent and severe form of cerebrovascular disease and a leading cause of significant neurological disability. As the primary cause of hospitalization for neurological disorders, it necessitates urgent and effective therapeutic interventions. Xijiao Dihuang decoction (XJDHT), a classical traditional Chinese medicinal formula, is traditionally known for its effects in heat clearing, detoxification, blood cooling, and blood stasis removing. Emerging evidence suggests its potential to promote neurological recovery following ischemic stroke; however, systematic evaluation remains limited. This study employed a systematic network pharmacology approach to elucidate the molecular mechanisms underlying XJDHT’s therapeutic effects on ischemic stroke. LC-MS identified 24 bioactive compounds in XJDHT, using Traditional Chinese Medicine Systems Pharmacology and SwissTargetPrediction databases for target prediction. Disease targets for ischemic stroke were retrieved from GeneCards, Online Mendelian Inheritance in Man, Therapeutic Target Database, and Gene Expression Omnibus datasets. Intersection analysis found 424 overlapping targets between 690 compound targets and 3415 disease targets. Cytoscape and STRING platforms were used to construct networks and identify hub genes and critical modules. Functional enrichment analysis was conducted using the Database for Annotation, Visualization, and Integrated Discovery database. Network analysis identified core bioactive constituents, including paeoniflorin, albiflorin, benzoylpaeoniflorin, kaempferol, paeonol, ellagic acid, pinostrobin, β-sitosterol, and stigmasterol. Key therapeutic targets comprised phosphatidylinositol-4,5-bisphosphate 3-kinase regulatory subunit 1, SRC proto-oncogene, signal transducer and activator of transcription 3, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit beta, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit delta, AKT serine/threonine kinase 1, epidermal growth factor receptor, estrogen receptor 1, Harvey rat sarcoma virus, and neuroblastoma RAS viral oncogene homolog. The Kyoto Encyclopedia of Genes and Genomes pathway enrichment revealed significant involvement in the mammalian target of rapamycin signaling pathway, mitogen-activated protein kinase cascade, phosphatidylinositol 3-Kinase-Protein Kinase B (PI3K-AKT) pathway, erythroblastic leukemia viral oncogene homolog signaling, and vascular endothelial growth factor-mediated angiogenesis. These findings suggest XJDHT exerts neuroprotective effects through coordinated modulation of inflammatory responses and cellular signaling cascades, exhibiting the characteristic therapeutic properties of a multicomponent, multitarget, and multipathway approach.