基于网络药理学及体内实验探讨射麻止喘液治疗中性粒细胞型哮喘的分子机制  

Study on the Molecular Mechanism of Shema Zhichuan Liquid in the Treatment of Neutrophilic Asthma Based on Network Pharmacology and In Vivo Experiment

作　　者：连乐燊[1,2] 孟星汝 黄秀芳 周谨希 谢燕小 陶海澜 蒋紫云[1,5] 刘小虹 LIAN Leshen;MENG Xingru;HUANG Xiufang;ZHOU Jinxi;XIE Yanxiao;TAO Hailan;JIANG Ziyun;LIU Xiaohong(Dongguan Hospital Affiliated to Guangzhou University of Chinese Medicine,Dongguan 523000 Guangdong,China;The First Clinical Medical School of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;Lingnan Medical Research Center of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;The Third Clinical Medical School of Guangzhou University of Chinese Medicine,Guangzhou 510370 Guangdong,China)

机构地区：[1]广州中医药大学东莞医院,广东东莞523000 [2]广州中医药大学第一临床医学院,广东广州510405 [3]广州中医药大学第一附属医院,广东广州510405 [4]广州中医药大学岭南医学研究中心,广东广州510405 [5]广州中医药大学第三临床医学院,广东广州510370

出　　处：《中药新药与临床药理》2024年第2期247-256,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省基础与应用基础研究基金项目(2023A1515010807,2021A1515010146);刘小虹广东省名中医传承工作室建设项目(粤中医办函[2018]5号);广州中医药大学东莞医院拔尖型临床人才培养项目(东中医[2022]158号);深圳市“医疗卫生三名工程”建设项目(SZZYSM202206013)。

摘　　要：目的基于网络药理学及体内实验探讨射麻止喘液治疗中性粒细胞型哮喘(NA)的分子机制。方法(1)利用中药系统药理学数据库和分析平台(TCMSP)、文献检索及Swiss ADME、Swiss Target Prediction数据库检索、筛选射麻止喘液的活性成分及其作用靶点。利用OMIM、Gene Cards、Dis Ge NET及Drug Bank数据库检索、筛选NA疾病相关靶点。通过微生信平台对射麻止喘液活性成分作用靶点与NA疾病相关靶点取交集,得到射麻止喘液治疗NA的潜在作用靶点(共有靶点)。采用Cytoscape 3.8软件构建“中药-活性成分-潜在作用靶点”网络。通过STRING数据库建立潜在作用靶点蛋白互作(PPI)网络,使用内置Mcode插件分析获得核心靶点。使用Metascape平台对潜在作用靶点进行基因本体(GO)、京都基因和基因组百科全书(KEGG)通路富集分析。(2)将BALB/C小鼠适应性喂养1周后,随机分为空白组、NA模型组、射麻止喘液低剂量组(2.5 g·kg^(-1))、射麻止喘液高剂量组(10 g·kg^(-1))、地塞米松对照组(1 mg·kg^(-1));通过腹腔注射致敏剂[鸡卵清白蛋白(OVA)+弗氏完全佐剂(CFA)]并通过雾化吸入激发的方法复制NA小鼠模型,第0、7、14天分别腹腔注射OVA+CFA(20μg OVA+75μg CFA,0.3 m L)致敏,第21~30天采用5%OVA混悬液雾化激发(每次8 m L,每次雾化时间40 min,每天1次);雾化激发前1 h各组灌胃给药,地塞米松对照组腹腔注射给药,每天1次。采用HE染色法观察小鼠肺组织病理变化;ELISA法检测肺泡灌洗液中IL-8含量;RT-q PCR法检测肺组织中NLRP3、CXCR2 m RNA表达水平;免疫组化法检测肺组织中p-m TOR蛋白表达水平。结果(1)共筛选得到射麻止喘液活性成分作用靶点826个,NA疾病相关靶点154个,取交集后得到射麻止喘液治疗NA的51个潜在作用靶点(共有靶点)。通过“中药-活性成分-潜在作用靶点”网络分析得到槲皮素、木犀草素、山柰酚、豆甾醇、柚皮素等关键活性成分Objective To investigate the molecular mechanism of Shema Zhichuan Liquid in the treatment of neutrophilic asthma(NA)based on network pharmacology and in vivo experiments.Methods(1)The TCMSP,literature search and Swiss ADME and Swiss Target Prediction databases were used to search and screen the active components and their targets of Shema Zhichuan Liquid.OMIM,GeneCards,DisGeNET and DrugBank databases were used to search and screen NA disease-related targets.The intersection of the active components and NA disease-related targets of Shema Zhichuan Liquid was obtained through the microbiology platform to obtain the potential targets of Shema Zhichuan Liquid for the treatment of NA(common targets).Cytoscape 3.8 software was used to construct the network of"Chinese medicinals-active components-potential targets".The PPI network of potential targets was established by STRING database,and the core targets were obtained by analysing the built-in Mcode plug-in.The Metascape platform was used to enrich the gene ontology(GO),Kyoto Encyclopaedia of Genes and Genomes(KEGG)pathways for the potential targets.(2)BALB/C mice were acclimatised and fed for 1 week and randomly divided into a blank group,NA model group,low-dose group(2.5 g·kg^(-1))and high-dose group of Shema Zhichuan Liquid(10 g·kg^(-1)),and control group of Dexamethasone(1 mg·kg^(-1));the NA mouse model was replicated by intraperitoneal injection of sensitizer(OVA+CFA)and nebulized inhalation excitation.OVA/CFA(20μg OVA+75μg CFA,0.3 mL)was injected intraperitoneally to sensitize on days 0,7 and 14 respectively,and 5%OVA suspension was nebulized on days 21-30(8 mL each time,40 minutes each time,once a day);1 hour before nebulisation,each group was administered by gastric gavage,and the Dexamethasone control group was administered by intraperitoneal injection once a day.The pathological changes of mouse lung tissue were observed by HE staining;IL-8 content in alveolar lavage fluid was detected by ELISA;mRNA expression levels of NLRP3 and CXCR2 were detected by

关 键 词：射麻止喘液 中性粒细胞型哮喘 网络药理学 NLRP3 CXCR2 MTOR信号通路 小鼠 

分 类 号：R285.5[医药卫生—中药学] R857.3[医药卫生—中医学]