目的 將金銀花化學(xué)成分與網(wǎng)絡(luò)藥理學(xué)研究結(jié)合，探討金銀花抗新型冠狀病毒肺炎（COVID-19）的潛在作用機制。方法 采用四極桿-靜電場軌道阱高分辨質(zhì)譜技術(shù)（UPLC-Q-Exactive/MS）對金銀花化學(xué)成分進行定性分析，根據(jù)色譜峰保留時間、精確相對分子質(zhì)量、特征碎片離子，結(jié)合相關(guān)文獻數(shù)據(jù)，鑒定金銀花的化學(xué)成分；通過SWISSTargetPrediction數(shù)據(jù)庫獲得成分靶點，通過GeneCards和OMIM數(shù)據(jù)庫獲得COVID-19疾病靶點，利用Venny2.1.0獲得金銀花和COVID-19共同的藥效靶點，利用Cytoscape 3.7.2軟件構(gòu)建“成分-靶點”網(wǎng)絡(luò)關(guān)系圖；以STRING數(shù)據(jù)庫構(gòu)建蛋白質(zhì)-蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)關(guān)系圖并對靶點進行基因本體（GO）功能富集分析和京都基因與基因組百科全書（KEGG）通路富集分析；利用分子對接技術(shù)驗證核心靶點與相對應(yīng)的成分的結(jié)合能力；利用體外抗病毒檢測驗證金銀花中活性成分綠原酸抗COVID-19的潛在作用機制。結(jié)果 共鑒定出金銀花化學(xué)成分77個。咖啡酸、腺苷、5,3'-二甲氧基木犀草素、芹菜素、阿魏酸等5個成分可能是金銀花抗COVID-19的活性成分，其中表皮生長因子受體（EGFR）、白細(xì)胞介素-2（IL-2）為關(guān)鍵靶點，涉及低氧誘導(dǎo)因子-1信號通路（HIF-1 signaling pathway）、磷脂酰肌醇-3-激酶-蛋白激酶B信號通路（PI3K-Akt signaling pathway）等。進一步分子對接結(jié)果表明金銀花的5個關(guān)鍵活性成分與疾病的2個核心靶點均具有良好的結(jié)合活性，結(jié)合能均＜-20.9 kJ·mol^-1。同時金銀花中主要有效成分綠原酸對3CLpro具有較好的體外抑制活性［半數(shù)抑制濃度（IC₅₀）為（106.40±0.71） μmol·L^-1］。結(jié)論 金銀花可能通過EGFR、IL2和3CLpro等多靶點發(fā)揮抗COVID-19作用，可進一步探討金銀花抗COVID-19的作用機制。

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

國家自然科學(xué)基金資助項目(82274397);“新高校20條”資助項目(自主培養(yǎng)創(chuàng)新團隊)(2021GXRC028);國家中醫(yī)藥管理局高水平中醫(yī)藥重點學(xué)科建設(shè)項目(ZYYZDXK-2023119);山東省自然基金項目(中醫(yī)藥聯(lián)合基金)(ZR2021LZY012);上海市復(fù)方中藥重點實驗室(上海中醫(yī)藥大學(xué))開放課題基金資助(21DZ2270500);2023年度濟南市市校融合發(fā)展戰(zhàn)略工程項目(JN-SX2023054);山東省技術(shù)創(chuàng)新引導(dǎo)計劃(YDZX2023044)