目的 将金银花化学成分与网络药理学研究结合，探讨金银花抗新型冠状病毒肺炎（COVID-19）的潜在作用机制。方法 采用四极杆-静电场轨道阱高分辨质谱技术（UPLC-Q-Exactive/MS）对金银花化学成分进行定性分析，根据色谱峰保留时间、精确相对分子质量、特征碎片离子，结合相关文献数据，鉴定金银花的化学成分；通过SWISSTargetPrediction数据库获得成分靶点，通过GeneCards和OMIM数据库获得COVID-19疾病靶点，利用Venny2.1.0获得金银花和COVID-19共同的药效靶点，利用Cytoscape 3.7.2软件构建“成分-靶点”网络关系图；以STRING数据库构建蛋白质-蛋白质相互作用（PPI）网络关系图并对靶点进行基因本体（GO）功能富集分析和京都基因与基因组百科全书（KEGG）通路富集分析；利用分子对接技术验证核心靶点与相对应的成分的结合能力；利用体外抗病毒检测验证金银花中活性成分绿原酸抗COVID-19的潜在作用机制。结果 共鉴定出金银花化学成分77个。咖啡酸、腺苷、5,3'-二甲氧基木犀草素、芹菜素、阿魏酸等5个成分可能是金银花抗COVID-19的活性成分，其中表皮生长因子受体（EGFR）、白细胞介素-2（IL-2）为关键靶点，涉及低氧诱导因子-1信号通路（HIF-1 signaling pathway）、磷脂酰肌醇-3-激酶-蛋白激酶B信号通路（PI3K-Akt signaling pathway）等。进一步分子对接结果表明金银花的5个关键活性成分与疾病的2个核心靶点均具有良好的结合活性，结合能均＜-20.9 kJ·mol^-1。同时金银花中主要有效成分绿原酸对3CLpro具有较好的体外抑制活性［半数抑制浓度（IC₅₀）为（106.40±0.71） μmol·L^-1］。结论 金银花可能通过EGFR、IL2和3CLpro等多靶点发挥抗COVID-19作用，可进一步探讨金银花抗COVID-19的作用机制。

Objective To explore the potential mechanism of Lonicerae Japonicae Flos against COVID-19 by combining the chemical composition of Lonicerae Japonicae Flos with network pharmacology study. Methods The chemical constituents of Lonicerae Japonicae Flos were qualitatively analyzed by UPLC-Q-Exactive/MS. The chemical constituents of Lonicerae Japonicae Flos were identified according to the peak retention time, exact molecular mass, characteristic fragment ions, and related literature data. The targets of constituents were obtained by SWISS Target Prediction database, the disease targets of COVID-19 were obtained by GeneCard and OMIM database, the common pharmacodynamic targets of Lonicerae Japonicae Flos and COVID-19 were obtained by Venny 2.1.0, and the "constituent-target" network was constructed by Cytoscape 3.7.2 software. The protein-protein interaction (PPI) network was constructed by STRING database, and the GO analysis and KEGG analysis were performed on the targets. The molecular docking technology was used to verify the binding ability of core targets and corresponding constituents. The potential mechanism of chlorogenic acid in Lonicerae Japonicae Flos against COVID-19 was verified by in vitro antiviral test. Results A total of 77 chemical constituents were identified from Lonicerae Japonicae Flos in vitro. Caffeic acid, adenosine, chrysoeriol 5-methyl ether, apigenin and ferulic acid, may be the active components of Lonicerae Japonicae Flos against COVID-19. Epidermal growth factor receptor (EGFR) and interleukin-2 (IL2) were the key targets, involving hypoxia inducible factor-1 (HIF-1) signaling pathway, phosphatidylinositol 3-kinase-protein kinase B (PI3K-AKT) signaling pathway. Further molecular docking results showed that the five key active components of Lonicerae Japonicae Flos and the two core targets of the disease had good binding activity, and the binding energy values were less than -20.9 kJ·mol^-1. At the same time, chlorogenic acid, the main effective component of Lonicerae Japonicae Flos, had good in vitro inhibitory activity against 3CLpro [IC₅₀ was (106.40 ±0.71) μmol·L^-1]. Conclusions Lonicerae Japonicae Flos may inhibit COVID-19 through multiple targets such as EGFR, IL2 and 3CLpro, which can provide a valuable basis for further research on the mechanism of Lonicerae Japonicae Flos against COVID-19.

R285.5

国家自然科学基金资助项目(82274397);“新高校20条”资助项目(自主培养创新团队)(2021GXRC028);国家中医药管理局高水平中医药重点学科建设项目(ZYYZDXK-2023119);山东省自然基金项目(中医药联合基金)(ZR2021LZY012);上海市复方中药重点实验室(上海中医药大学)开放课题基金资助(21DZ2270500);2023年度济南市市校融合发展战略工程项目(JN-SX2023054);山东省技术创新引导计划(YDZX2023044)