目的 研究梔子抗呼吸道合胞病毒（RSV）作用并預(yù)測其機制。方法 通過細胞毒性實驗、細胞病變效應(yīng)（CPE）及實時熒光定量PCR（qRT-PCR）實驗，探索梔子提取物對RSV的抑制作用；在中藥系統(tǒng)藥理學(xué)分析平臺（TCMSP）中搜索梔子的活性成分并預(yù)測其靶點，在GeneCards、OMIM、Disgen數(shù)據(jù)庫中獲取RSV相關(guān)靶點，獲取藥物-病毒交集基因后構(gòu)建關(guān)鍵蛋白質(zhì)-蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)，并用分子對接進行驗證；交集基因通過DAVID數(shù)據(jù)庫進行基因本體（GO）功能富集分析和基于京都基因與基因組百科全書（KEGG）通路富集分析，預(yù)測其作用機制。結(jié)果 梔子提取物半數(shù)細胞毒性濃度（TC₅₀）為4.621 mg·mL^-1，質(zhì)量濃度為3.543 8、1.771 9、0.886 0 mg·mL^-1時有顯著抗RSV作用，且隨質(zhì)量濃度增加，抗RSV作用越強。網(wǎng)絡(luò)藥理學(xué)共篩選到梔子有效化合物10個［口服生物利用度（OB）≥30%，類藥性（DL）＞0.18），可視化結(jié)果顯示有118個節(jié)點，主要靶點有蛋白激酶Bα （AKT1）、非受體酪氨酸激酶（SRC）、表皮生長因子（EGFR）等；GO結(jié)果顯示主要與蛋白激酶B信號的正向調(diào)節(jié)、蛋白質(zhì)磷酸化等生物過程（BP）；質(zhì)膜、大分子復(fù)合物等細胞組成（CC）； ATP結(jié)合、酶結(jié)合、蛋白激酶活性等分子功能（MF）有關(guān)。KEGG通路富集到117條信號通路（FDR＜0.01），主要與腫瘤中程序性死亡分子1配體（PD-L1）的表達和程序性死亡分子1（PD-1）檢查點通路、磷脂酰肌醇3-激酶/蛋白激酶B（PI3K-Akt）、C型凝集素受體（CLRs）等信號通路有關(guān)。結(jié)論 梔子提取物有顯著抗RSV作用，可能主要以豆甾醇、吡嗪環(huán)辛烯-4a-羧酸、β-谷甾醇、藏紅花酸等成分與EGFR、SRC、絲裂原活化蛋白激酶3（MAPK3）等靶點相結(jié)合，通過PD-L1表達和PD-1、PI3K-Akt、CLRs通路等信號通路發(fā)揮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.

R965

貴州省十大工業(yè)（健康醫(yī)藥）產(chǎn)業(yè)振興專項資金資助（黔財工［2020］247號）;貴州省中醫(yī)藥管理局中醫(yī)藥、民族醫(yī)藥科學(xué)技術(shù)研究專項課題（QZYY-2024-188）;貴州中醫(yī)藥大學(xué)研究生教育創(chuàng)新計劃資助項目（YCXKYS2023034）