目的 研究栀子抗呼吸道合胞病毒（RSV）作用并预测其机制。方法 通过细胞毒性实验、细胞病变效应（CPE）及实时荧光定量PCR（qRT-PCR）实验，探索栀子提取物对RSV的抑制作用；在中药系统药理学分析平台（TCMSP）中搜索栀子的活性成分并预测其靶点，在GeneCards、OMIM、Disgen数据库中获取RSV相关靶点，获取药物-病毒交集基因后构建关键蛋白质-蛋白质相互作用（PPI）网络，并用分子对接进行验证；交集基因通过DAVID数据库进行基因本体（GO）功能富集分析和基于京都基因与基因组百科全书（KEGG）通路富集分析，预测其作用机制。结果 栀子提取物半数细胞毒性浓度（TC₅₀）为4.621 mg·mL^-1，质量浓度为3.543 8、1.771 9、0.886 0 mg·mL^-1时有显著抗RSV作用，且随质量浓度增加，抗RSV作用越强。网络药理学共筛选到栀子有效化合物10个［口服生物利用度（OB）≥30%，类药性（DL）＞0.18），可视化结果显示有118个节点，主要靶点有蛋白激酶Bα （AKT1）、非受体酪氨酸激酶（SRC）、表皮生长因子（EGFR）等；GO结果显示主要与蛋白激酶B信号的正向调节、蛋白质磷酸化等生物过程（BP）；质膜、大分子复合物等细胞组成（CC）； ATP结合、酶结合、蛋白激酶活性等分子功能（MF）有关。KEGG通路富集到117条信号通路（FDR＜0.01），主要与肿瘤中程序性死亡分子1配体（PD-L1）的表达和程序性死亡分子1（PD-1）检查点通路、磷脂酰肌醇3-激酶/蛋白激酶B（PI3K-Akt）、C型凝集素受体（CLRs）等信号通路有关。结论 栀子提取物有显著抗RSV作用，可能主要以豆甾醇、吡嗪环辛烯-4a-羧酸、β-谷甾醇、藏红花酸等成分与EGFR、SRC、丝裂原活化蛋白激酶3（MAPK3）等靶点相结合，通过PD-L1表达和PD-1、PI3K-Akt、CLRs通路等信号通路发挥抗呼吸道合胞病毒作用。

Objective To study the anti-respiratory syncytial virus (RSV) effect of Gardeniae Fructus and predict its mechanism. Methods To explore the inhibitory effect of Gardeniae Fructus on respiratory syncytial virus (RSV) through drug cytotoxicity assay, cytopathic effect (CPE) and real-time fluorescence quantitative PCR (qRT-PCR) assay; to search for the active ingredients of Gardeniae Fructus and predict its targets in TCMSP, and to obtain the relevant targets of RSV from GeneCards, OMIM and Disgen databases; to construct key protein-protein interactions after obtaining drug-virus intersection genes. RSV related targets were obtained from GeneCards, OMIM, and Disgen databases, and the drug-virus intersection genes were obtained to construct a key protein-protein interaction (PPI) network and validated by molecular docking; the intersection genes were analyzed by Gene Ontology Functional Enrichment (GO) analysis via the DAVID database, and the mechanism of action was predicted by the Kyoto Encyclopedia of Genes and Genomes (KEGG) -based pathway enrichment analysis. Results The half cytotoxicity concentration (TC₅₀) of Gardeniae Fructus extract was 4.621 mg·mL^-1, and significant anti-RSV effects were observed at mass concentrations of 3.543 8, 1.771 9, and 0.886 0 mg·mL^-1, and with the increase of the mass concentration, the anti-RSV effects were stronger. A total of 10 effective compounds of Gardeniae Fructus were screened by network pharmacology (OB ≥ 30%, DL > 0.18), and the visualization results showed that there were 118 nodes, and the main targets were AKT1, SRC, and EGFR, etc.; GO results showed that they were mainly related to the processes of positive regulation of protein kinase B signaling, and protein phosphorylation; the plasma membrane, and the macromolecule complex; and the processes of ATP-binding, enzyme-binding, and protein kinase activity. processes. KEGG pathway was enriched to 117 signaling pathways (FDR < 0.01), which were mainly related to PD-L1 expression and PD-1 checkpoint pathway, phosphatidylinositol 3-kinase/protein kinase B (PI3K-Akt), and C-type lectin receptors (CLRs) signaling pathways in tumors. Conclusion Gardeniae Fructus extract has significant anti-RSV effects, which may be mainly based on the combination of components such as soy sterol, β-glutamate, and saffronic acid with targets such as EGFR, SRC, and MAPK3, and exerts its anti-respiratory syncytial virus effects through the signaling pathways such as PD-L1 expression and the PD-1, PI3KAkt, and CLRs pathways.

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贵州省十大工业（健康医药）产业振兴专项资金资助（黔财工［2020］247号）;贵州省中医药管理局中医药、民族医药科学技术研究专项课题（QZYY-2024-188）;贵州中医药大学研究生教育创新计划资助项目（YCXKYS2023034）