目的 采用UPLC-QTOF/MS技術(shù)與網(wǎng)絡(luò)藥理學(xué)方法探究草烏抗大腸埃希菌的作用機制。方法 采用UPLC-QTOF/MS技術(shù)鑒定草烏在大鼠血清中的主要成分，并結(jié)合網(wǎng)絡(luò)藥理學(xué)識別其作用靶點。利用Cytoscape和STRING數(shù)據(jù)庫構(gòu)建“成分-靶點-疾病”網(wǎng)絡(luò)和蛋白相互作用（PPI）網(wǎng)絡(luò)圖，進一步通過基因本體（GO）和京都基因與基因組百科全書（KEGG）通路分析揭示草烏抗大腸埃希菌的潛在作用機制。通過分子模擬技術(shù)評估關(guān)鍵入血成分與靶標(biāo)蛋白的結(jié)合效力。結(jié)果 鑒定出草烏中的152個化合物和40個入血成分，其中8個為原型成分，32個為代謝產(chǎn)物。確定大腸埃希菌的928個相關(guān)靶點，并通過韋恩圖識別出90個草烏作用的交集靶點。PPI網(wǎng)絡(luò)分析顯示草烏入血成分主要作用于原癌基因酪氨酸蛋白激酶（SRC）、B淋巴細胞瘤-2（Bcl-2）、表皮生長因子受體（EGFR）、雌激素受體1（ESR1）、V-Jun肉瘤病毒癌基因同源物（JUN）、胱天蛋白酶3（CASP3）等關(guān)鍵蛋白。GO功能富集分析表明草烏的作用機制主要與生物學(xué)過程中的激素反應(yīng)、有機環(huán)化合物反應(yīng)、脂質(zhì)反應(yīng)等相關(guān)；在細胞定位中與膜筏、膜微域、質(zhì)膜筏等有關(guān)；分子功能則涉及肽酶類的多種活性。KEGG通路分析揭示草烏可能通過焦點黏附、白細胞跨內(nèi)皮遷移、自然殺傷細胞介導(dǎo)的細胞毒性、Rap1信號通路、細胞間隙連接、中性粒細胞胞外誘捕網(wǎng)等途徑發(fā)揮作用。分子對接結(jié)果顯示，?；悄懰帷⑸n術(shù)素、吳茱萸新堿等成分與Bcl-2、EGFR、ESR1等關(guān)鍵蛋白有良好的結(jié)合。結(jié)論 結(jié)合UPLC-QTOF/MS技術(shù)、網(wǎng)絡(luò)藥理學(xué)分析和分子對接模擬，推測了蒙藥草烏抗大腸埃希菌的作用機制，為其抗菌作用的進一步研究和應(yīng)用提供科學(xué)依據(jù)。

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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內(nèi)蒙古自治區(qū)自然科學(xué)基金項目(2021LHMS08055);內(nèi)蒙古醫(yī)科大學(xué)科技創(chuàng)新團隊(RZ2200002685);內(nèi)蒙古醫(yī)科大學(xué)蒙藥學(xué)“一流學(xué)科”青年教師創(chuàng)新能力提升項目(myxy1xkky2020-02);內(nèi)蒙古醫(yī)科大學(xué)蒙藥學(xué)“一流學(xué)科”研究生科研能力提升計劃項目(MYX2023-R03)