目的 采用UPLC-QTOF/MS技术与网络药理学方法探究草乌抗大肠埃希菌的作用机制。方法 采用UPLC-QTOF/MS技术鉴定草乌在大鼠血清中的主要成分，并结合网络药理学识别其作用靶点。利用Cytoscape和STRING数据库构建“成分-靶点-疾病”网络和蛋白相互作用（PPI）网络图，进一步通过基因本体（GO）和京都基因与基因组百科全书（KEGG）通路分析揭示草乌抗大肠埃希菌的潜在作用机制。通过分子模拟技术评估关键入血成分与靶标蛋白的结合效力。结果 鉴定出草乌中的152个化合物和40个入血成分，其中8个为原型成分，32个为代谢产物。确定大肠埃希菌的928个相关靶点，并通过韦恩图识别出90个草乌作用的交集靶点。PPI网络分析显示草乌入血成分主要作用于原癌基因酪氨酸蛋白激酶（SRC）、B淋巴细胞瘤-2（Bcl-2）、表皮生长因子受体（EGFR）、雌激素受体1（ESR1）、V-Jun肉瘤病毒癌基因同源物（JUN）、胱天蛋白酶3（CASP3）等关键蛋白。GO功能富集分析表明草乌的作用机制主要与生物学过程中的激素反应、有机环化合物反应、脂质反应等相关；在细胞定位中与膜筏、膜微域、质膜筏等有关；分子功能则涉及肽酶类的多种活性。KEGG通路分析揭示草乌可能通过焦点黏附、白细胞跨内皮迁移、自然杀伤细胞介导的细胞毒性、Rap1信号通路、细胞间隙连接、中性粒细胞胞外诱捕网等途径发挥作用。分子对接结果显示，牛磺胆酸、苍术素、吴茱萸新碱等成分与Bcl-2、EGFR、ESR1等关键蛋白有良好的结合。结论 结合UPLC-QTOF/MS技术、网络药理学分析和分子对接模拟，推测了蒙药草乌抗大肠埃希菌的作用机制，为其抗菌作用的进一步研究和应用提供科学依据。

Objective To elucidate the mechanism of action of the Aconiti Kusnezoffii Radix against Escherichia coli, employing advanced analytical and computational methods. Methods Utilizing UPLC-QTOF/MS, the main components in rat serum were identified. Network pharmacology approaches were used to identify the action targets. Cytoscape and STRING databases were employed to construct “component - target - disease” networks and protein-interaction maps. GO and KEGG pathway analyses were conducted to explore the potential mechanisms of action. Molecular modeling techniques were applied to assess the binding affinity of key blood entry components to target proteins. Results Aconiti Kusnezoffii Radix was found to contain 152 compounds and 40 blood-entry components, including 8 prototypical components and 32 metabolites. The study identified 928 relevant targets of Escherichia coli, with 90 intersecting targets of Aconiti Kusnezoffii Radix’s action identified through the Venn diagram. PPI network analysis revealed that the blood-entry components primarily interacted with key proteins such as SRC, Bcl-2, EGFR, ESR1, JUN, and CASP3. GO functional enrichment analysis showed that the mechanism of action was mainly related to hormonal reaction, organic cyclic compound reaction and lipid reaction in biological processes. In cell localization, it is related to membrane raft, membrane microdomain, plasma membrane raft, etc. Molecular functions involve various activities of peptidases. KEGG pathway analysis suggested potential mechanisms such as focal adhesion, leukocyte transendothelial migration, natural killer cell-mediated cytotoxicity, Rap1 signaling pathway, cell gap junctions, and the extracellular trapping network of neutrophils. Molecular docking analysis showed that compounds like taurocholic acid, atractylodin, and evocarpine have a strong binding affinity to key proteins such as Bcl-2, EGFR, and ESR1. Conclusion Combined with UPLC-QTOF/MS technique, network pharmacological analysis and molecular docking simulation, the mechanism of action of Aconiti Kusnezoffii Radix against Escherichia coli, which provided scientific basis for further research and application of its antibacterial action.

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内蒙古自治区自然科学基金项目(2021LHMS08055);内蒙古医科大学科技创新团队(RZ2200002685);内蒙古医科大学蒙药学“一流学科”青年教师创新能力提升项目(myxy1xkky2020-02);内蒙古医科大学蒙药学“一流学科”研究生科研能力提升计划项目(MYX2023-R03)