病毒学报

目的 系统评价金振口服液（Jinzhen oral liquid, JZOL）对甲型H1N1流感病毒PR8株（H1N1/PR8）及呼吸道合胞病毒（Respiratory syncytial virus, RSV）感染所致肺炎小鼠模型的治疗作用，通过网络药理学方法研究JZOL治疗H1N1与RSV感染致肺炎的作用机制，为JZOL的临床应用提供实验药理学依据。方法 建立H1N1/PR8病毒感染的小鼠肺炎模型，观察JZOL对小鼠肺指数、肺组织病毒载量和死亡保护的作用。建立RSV病毒感染的小鼠肺炎模型，考察JZOL对小鼠肺指数及肺指数抑制率的影响。利用中药系统药理学数据库和分析平台筛选JZOL中药物的活性成分与作用靶点，通过GeneCards数据库检索H1N1与RSV相关靶点，统一利用Uniprot数据库进行标准化，采用Cytoscape 3.10.4软件建立JZOL成分-靶点网络图，借助Venny 2.1.0在线平台获取JZOL药物靶点分别与H1N1和RSV疾病靶点的交集靶点，将交集靶点上传至STRING 12.0数据库获得蛋白质-蛋白质相互作用（PPI）关系网络，并在Metascape平台中进行基因本体（GO）富集分析和京都基因与基因组百科全书（KEGG）分析。结果 JZOL3个剂量组均可不同程度的降低H1N1/PR8感染小鼠和RSV病毒感染小鼠肺指数，在H1N1/PR8感染模型中，JZOL还能有效降低肺组织病毒载量，并对感染小鼠表现出明确的死亡保护作用。通过TCMSP平台筛选出JZOL主要活性成分141个，对应作用靶点238个，JZOL-H1N1交集靶点61个，JZOL-RSV交集靶点112个，PPI网络分析提示，JZOL治疗H1N1可能关键作用于AKT1、IL-6、TNF、IL1B、TP53等靶点；治疗RSV则可能涉及IL-6、TNF、AKT1、IL1B、JUN等核心靶标。GO与KEGG富集分析表明，JZOL可能通过调控炎症反应、细胞凋亡、免疫应答等生物学过程，并作用于AGE-RAGE、IL-17、TNF、流感病毒感染、PI3K-Akt等多条信号通路，从而发挥治疗作用。结论 研究结果表明JZOL对H1N1/PR8流感病毒和RSV病毒感染肺炎小鼠具有显著治疗效果，其治疗作用通过多成分、多靶点、多通路发挥。

Objective To systematically evaluate the therapeutic effects of Jinzhen Oral Liquid(JZOL) on mouse pneumonia models induced by Influenza A(H1N1) PR8 virus(H1N1/PR8) and respiratory syncytial virus(RSV), and to investigate the underlying mechanisms of JZOL in the treatment of H1N1-and RSVinduced pneumonia using network pharmacology, thereby providing experimental pharmacological evidence for its clinical application.Methods A mouse pneumonia model was established by infection with the H1N1/PR8 virus to assess the effects of JZOL on lung index, pulmonary viral load, and survival protection. An RSVinduced mouse pneumonia model was also established to evaluate the effects of JZOL on lung index and lung index inhibition rate. For mechanism exploration, active components and corresponding targets of JZOL were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). H1N1-and RSV-related targets were retrieved from the GeneCards database and standardized using the UniProt database. A compound – target network of JZOL was constructed using Cytoscape 3.10.4. Overlapping targets between JZOL and H1N1 or RSV were identified using the Venny 2.1.0 platform. Protein–protein interaction(PPI) networks were generated using the STRING 12.0 database, followed by Gene Ontology(GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analyses using the Metascape platform.Results All three doses of JZOL significantly reduced the lung index in both H1N1/PR8-and RSV-infected mice. In the H1N1/PR8 infection model, JZOL also effectively decreased pulmonary viral load and provided a clear protective effect against mortality. Network pharmacology analysis identified 141 major active components of JZOL corresponding to 238 potential targets. A total of 61 overlapping targets were identified between JZOL and H1N1, and 112 overlapping targets between JZOL and RSV. PPI network analysis indicated that the therapeutic effects of JZOL against H1N1 infection may primarily involve key targets such as AKT1, IL-6, TNF, IL1B, and TP53, while its effects against RSV infection may be associated with core targets including IL-6, TNF, AKT1, IL1B, and JUN. GO and KEGG enrichment analyses suggested that JZOL exerts its therapeutic effects by regulating biological processes related to inflammation, apoptosis, and immune responses, and by modulating multiple signaling pathways, including AGE – RAGE, IL-17, TNF, influenza A, and PI3K – Akt signaling pathways. Conclusion Jinzhen Oral Liquid exhibits significant therapeutic effects on mouse pneumonia induced by H1N1/PR8 influenza virus and RSV. Its pharmacological activity is mediated through a multi-component, multi-target, and multi-pathway mechanism, providing a scientific basis for the clinical application of JZOL in the treatment of viral pneumonia.