摘要
目的基于网络药理学和动物实验验证探讨定喘颗粒干预呼吸道合胞病毒肺炎的作用机制。方法通过TCMSP、TCMID等数据库获取定喘颗粒中各个中药的潜在活性成分及作用靶点,根据ADME药动学参数筛选定喘颗粒中药物的活性成分;通过Genecards、OMIM、DisGeNet数据库获取呼吸道合胞病毒肺炎潜在的疾病靶点;利用String平台构建蛋白互作网络对交集靶点进行可视化处理;采用David数据库对关键核心靶点进行GO功能富集分析和KEGG通路富集分析;随后利用Cytoscape软件(3.9.1)构建定喘颗粒干预呼吸道合胞病毒肺炎的相关网络。选取呼吸道合胞病毒(RSV)致呼吸道合胞病毒肺炎幼龄大鼠模型进行实验验证。结果网络药理学结果显示定喘颗粒共177个潜在活性成分,作用于144个靶点,呼吸道合胞病毒肺炎共1075个相关靶点,得到“药物-疾病”交集靶点55个,其中核心靶点25个为ALB、IL6、CASP3、EGFR、VEGFA等;GO功能富集分析结果显示涉及的生物过程主要包括对转录的正调控、对RNA聚合酶Ⅱ启动子转录的正调控、对基因表达的正调控、对细胞凋亡过程的负调控等;KEGG通路富集主要涉及PI3KAkt信号通路、癌症通路、肿瘤坏死因子信号通路、IL-17信号通路等。动物实验初步表明:定喘颗粒可通过调控PI3K-Akt信号通路,从而参与炎症、免疫反应发挥作用。与正常组相比,模型组幼龄大鼠肺组织中p-PI3K/PI3K、p-Akt/Akt的比值显著升高(P<0.05),病毒载量显著升高(P<0.05),肺组织损伤病理评分显著增高(P<0.05),肺泡灌洗液(BALF)中炎症因子IL-6、TNF-α含量明显增高(Ρ<0.05)。与模型组相比,定喘颗粒各剂量组和阳性对照组幼龄大鼠肺组织中p-PI3K/PI3K、p-Akt/Akt蛋白表达降低(Ρ<0.05),病毒载量显著降低(P<0.05),肺组织损伤病理评分明显降低(P<0.05),BALF中炎症因子IL-6、TNF-α含量明显降低(Ρ<0.05)。结论研究揭示了定喘颗粒治疗呼吸道合胞病毒肺炎的多组分、多靶点、多通路的协同作用机制。经动物实验验证,RSV感染幼龄大鼠时很有可能激活了PI3K-Akt信号通路,引起炎症因子IL-6、TNF-α释放,定喘颗粒能够有效下调PI3K-Akt信号通路,抑制炎症因子IL-6、TNF-α释放从而达到达到抗炎的作用。
Objective To investigate the mechanism of action of Dingchuan granules in intervening respiratory syncytial virus pneumonia based on network pharmacology and animal experiments.Methods The potential active ingredients and targets of each traditional Chinese medicine in Dingchuan granules were obtained from TCMSP and TCMID databases,and the active ingredients of the drugs in Dingchuan granules were screened according to ADME pharmacokinetic parameters;the potential disease targets of respiratory syncytial virus pneumonia were obtained from Genecards,OMIM,and DisGeNet databases;the protein-interaction networks of intersecting targets were visualized by using String platform;the key core targets were visualized by using David database;and the intersection targets were visualized by using David database.Interaction networks were constructed using the String platform to visualize the intersecting targets;GO functional enrichment and KEGG pathway enrichment analyses of the key core targets were performed using the David database;and then the relevant networks for the intervention of Respiratory Syncytial Virus Pneumonia in the Dingchuan particles were constructed using the Cytoscape software(3.9.1).Respiratory syncytial virus(RSV)-induced respiratory syncytial virus pneumonia in young rats was selected for experimental validation.Results The results of the network pharmacology showed that the 177 potential active ingredients of Dingchuan granules acted on 144 targets,and there were 1075 targets related to respiratory syncytial virus pneumonia,and 55 drug-disease intersecting targets were obtained,of which 25 core targets were ALB,IL6,CASP3,EGFR,VEGFA,etc.The GO function of Dingchuan granules was also investigated,and the GO function of Dingchuan was investigated.The results of GO function enrichment analysis showed that the biological processes involved mainly include positive regulation of transcription,positive regulation of RNA polymerase II promoter transcription,positive regulation of gene expression,negative regulation of apoptosis,etc.The KEGG pathway enrichment mainly involves the PI3K-Akt signaling pathway,the cancer pathway,the tumor necrosis factor signaling pathway,the IL-17 signaling pathway and so on.The KEGG pathway enrichment mainly involves PI3K-Akt signaling pathway,tumor necrosis factor signaling pathway,IL-17 signaling pathway,etc.Animal experiments initially showed that the fixed wheezing granules can play a role in inflammation and immune response by regulating the PI3K-Akt signaling pathway,thus participating in the inflammatory and immune response.Compared with the normal group,the ratios of p-PI3K/PI3K and p-Akt/Akt in the lung tissues of young rats in the model group were significantly higher(P<0.05),the viral load was significantly higher(P<0.05),the pathological score of lung tissue damage was significantly higher(P<0.05),and the content of inflammatory factors IL-6 and TNF-α in alveolar lavage fluid(BALF)was significantly higher(P<0.05).Compared with the model group,the expression of p-PI3K/PI3K and p-Akt/Akt proteins in the lung tissues of young rats in each dose group and the positive control group was reduced(P<0.05),the viral load was significantly reduced(P<0.05),the pathological scores of lung tissue injury were significantly reduced(P<0.05),and the contents of inflammatory factors IL-6 and TNF-α in BALF were significantly reduced(P<0.05).Conclusion The study revealed the synergistic mechanism of multicomponent,multi-target,and multi-pathway action of Dingchuan granules for the treatment of respiratory syncytial virus pneumonia.It was verified by animal experiments that RSV infection in young rats probably activated the PI3K-Akt signaling pathway,which caused the release of inflammatory factors IL-6 and TNF-α.Dingchuan granules could effectively down-regulate the PI3K-Akt signaling pathway,inhibit the release of inflammatory factors IL-6 and TNF-α,and thus achieve the anti-inflammatory effect.
作者
魏晨浩
张秀英
王雪峰
王咏雪
杨梦菲
刘清
赵航宇
Wei Chenhao;Zhang Xiuying;Wang Xuefeng;Wang Yongxue;Yang Mengfei;Liu Qing;Zhao Hangyu(The First Clinical College of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;The First Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第9期2996-3010,共15页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金委员会面上项目(8197152087):从“络脉玄微府”理论探讨清肺通络开玄法调控HMGB1/RAGE轴改善RSV诱导肺炎性损伤的药效机制研究,负责人:张秀英。
关键词
定喘颗粒
网络药理学
RSV肺炎
信号通路
靶点
Dingchuan granule
Network pharmacology
RSV pneumonia
Signaling pathway
Target point
作者简介
通讯作者:张秀英,主任医师,硕士研究生导师,主要研究方向:中医药防治儿童疾病。
