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ORIGINAL RESEARCH ARTICLE

Mechanistic investigation of Xi Jiao Di Huang Tang for ischemic stroke: A network pharmacology and in vivo validation study

Qiuhua He1 ,  Fantao Song1 ,  Yujie Wang1 ,  Zhaoyao Chen1*
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1 Department of Neurology, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China
Advanced Neurology, 025100020 https://doi.org/10.36922/AN025100020
Received: 4 March 2025 | Revised: 23 May 2025 | Accepted: 9 June 2025 | Published online: 4 July 2025
© 2025 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

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. Xi Jiao Di Huang Tang (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.

Keywords
Ischemic stroke
Traditional Chinese medicine
Network pharmacology
Micromolecular protein
Anti-inflammation
Antioxidation
Funding
This research was partially supported by the Key Project of Medical Research of the Jiangsu Commission of Health (K2023009) and the Natural Science Foundation of Nanjing University of Chinese Medicine (XZR2023030). The financial support was used to cover experimental expenses.
Conflict of interest
Zhaoyao Chen is the Youth Editorial Board Member of this journal but was not involved in the editorial and peer-review process conducted for this paper, either directly or indirectly. Separately, other authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper.
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