Table2_Investigation into the potential mechanism and molecular targets of Fufang Xueshuantong capsule for the treatment of ischemic stroke based on network pharmacology and molecular docking.DOCX

Fufang Xueshuantong (FFXST) capsule is a traditional Chinese medicine (TCM) preparation used to activate blood circulation, resolve stasis, benefit qi, and nourish yin in clinical practice. However, its potential mechanism and molecular targets after ischemic stroke (IS) have not been investigated. The aim of this research was to investigate the molecular mechanisms of FFXST in the treatment of IS based on network pharmacology and molecular docking. We used the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) to collect candidate compounds of four herbs in FFXST; disease-related differential genes were screened using the Gene Expression Omnibus (GEO) database, and a compound–disease network was created using Cytoscape 3.8.2 software. The topological analysis of the protein–protein interaction (PPI) network was then created to determine the candidate targets of FFXST against IS. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted using the clusterProfiler package in R. The gene–pathway network of FFXST against IS was created to obtain the key target genes. Molecular docking was used to validate the core targets using AutoDock Vina 1.1.2. A total of 455 candidate compounds of FFXST and 18,544 disease-related differential genes were screened. Among them, FFXST targets for IS treatment had 67 active compounds and 10 targets in the PPI network related to STAT1, STAT3, and HIF1A. The biological processes of GO analysis included the regulation of reactive oxygen species metabolic process, cellular response to chemical stress, regulation of angiogenesis, regulation of vasculature development, positive regulation of cytokine production, and response to oxidative stress. The KEGG enrichment analysis showed that Kaposi sarcoma-associated herpesvirus infection, microRNAs in the cancer signaling pathway, Th17 cell differentiation, and HIF-1 signaling pathway were significantly enriched. The network pharmacology outcomes were further verified by molecular docking. We demonstrated that FFXST protection against IS may relate to the regulation of oxidative stress, immune inflammatory response, and angiogenesis through the relevant signaling pathways. Our study systematically illustrated the application of network pharmacology and molecular docking in evaluating characteristics of multi-component, multi-target, and multi-pathway of FFXST for IS.

复方血栓通（Fufang Xueshuantong, FFXST）胶囊是一种中药（traditional Chinese medicine, TCM）制剂，临床中常用于活血化瘀、益气养阴。然而，其在缺血性脑卒中（ischemic stroke, IS）治疗中的潜在作用机制与分子靶点尚未得到系统研究。本研究旨在基于网络药理学与分子对接技术，探究复方血栓通胶囊治疗缺血性脑卒中的分子机制。我们通过中药系统药理学数据库与分析平台（Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform, TCMSP）收集复方血栓通胶囊中四味药材的候选化合物；利用基因表达综合数据库（Gene Expression Omnibus, GEO）筛选疾病相关差异基因，并通过Cytoscape 3.8.2软件构建化合物-疾病网络。随后构建蛋白质相互作用（protein–protein interaction, PPI）网络并开展拓扑分析，以确定复方血栓通胶囊抗缺血性脑卒中的候选靶点。采用R语言中的clusterProfiler包进行基因本体（Gene Ontology, GO）富集分析与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）富集分析；构建复方血栓通胶囊抗缺血性脑卒中的基因-通路网络，以获取关键靶基因。使用AutoDock Vina 1.1.2进行分子对接，验证核心靶点。本研究共筛选得到复方血栓通胶囊的455个候选化合物以及18544个疾病相关差异基因。其中，用于缺血性脑卒中治疗的复方血栓通胶囊靶点涉及67个活性化合物，以及PPI网络中与STAT1、STAT3、HIF1A相关的10个靶点。GO分析的生物学过程包括：活性氧代谢过程调控、细胞对化学应激的响应、血管生成调控、血管发育调控、细胞因子产生的正调控，以及氧化应激响应。KEGG富集分析显示，卡波西肉瘤相关疱疹病毒感染、癌症信号通路中的microRNAs、Th17细胞分化以及HIF-1信号通路显著富集。网络药理学结果经分子对接进一步验证。本研究证实，复方血栓通胶囊对缺血性脑卒中的保护作用可能通过相关信号通路调控氧化应激、免疫炎症反应与血管生成实现。本研究系统阐明了网络药理学与分子对接技术在评估复方血栓通胶囊治疗缺血性脑卒中多成分、多靶点、多通路特性中的应用价值。