基于网络药理学方法、分子对接技术及动物实验验证探讨通塞颗粒治疗慢性阻塞性肺疾病急性加重期的作用机制  

Mechanism of Action of Tongsai Granules in the Treatment of Acute Exacerbation of COPD Based on Network Pharmacology,Molecular Docking Technology,and Experimental Validation in Rats

作　　者：付子坚 李建生[2,3] 田燕歌[2,3] 赵鹏 燕苗苗 芦晓帆 FU Zijian;LI Jiansheng;TIAN Yange;ZHAO Peng;YAN Miaomiao;LU Xiaofan(The Second Clinical Medical School,Henan University of Chinese Medicine,Zhengzhou Henan 450011,China;Henan Key Laboratory of the TCM Prevention and Treatment of Respiratory Diseases,Henan University of Chinese Medicine,Zhengzhou Henan 450046,China;The First Clinical Medical School,Henan University of Chinese Medicine,Zhengzhou Henan450046,China)

机构地区：[1]河南中医药大学第二临床医学院,河南郑州450011 [2]河南中医药大学呼吸系统疾病重点实验室,河南郑州450046 [3]河南中医药大学第一临床医学院,河南郑州450046

出　　处：《新中医》2024年第6期170-185,共16页New Chinese Medicine

基　　金：国家自然科学基金项目(81904165);国家中医药临床科研基金建设项目(2019JDZX069)。

摘　　要：目的:基于网络药理学方法、分子对接技术和动物实验验证探讨通塞颗粒治疗慢性阻塞性肺疾病急性加重期(AECOPD)的作用机制。方法:(1)预测分析。通过中药系统药理学分析平台筛选通塞颗粒的活性成分及其对应靶点;利用人类基因(Genecards)、在线人类孟德尔遗传数据系统(OMIM)等数据库检索AECOPD疾病基因;绘制韦恩图并获取药物-疾病共同靶点;构建通塞颗粒活性成分-靶点网络图和蛋白质相互作用网络图,根据网络关系筛选核心靶点;借助R语言软件进行基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)通路富集分析,使用AutoDockTool 1.5.6软件对通塞颗粒关键活性成分和核心靶点进行分子对接。(2)实验验证。随机将50只SD大鼠分为对照组、模型组、中药组、西药组、联合组各10只。除对照组外,其余各组均构建AECOPD大鼠模型,并于急性加重前2 d及其后4 d予药物灌胃治疗。治疗结束后检测各组大鼠的肺功能指标[0.3 s用力呼气容积(FEV_(0.3))、用力肺活量(FVC)、0.3 s用力呼气容积与用力肺活量的比值(FEV_(0.3)/FVC)、呼气流量峰值(PEF)],检测血清肿瘤坏死因子(TNF)-α、白细胞介素(IL)-17含量,通过苏木精-伊红染色法(HE)观察气道与肺组织的病理改变,采用免疫印迹法和酶联免疫吸附测定法初步验证网络药理学的预测结果。结果:通塞颗粒治疗AECOPD的关键活性成分为槲皮素、芹黄素、木犀草素、山奈酚、β-谷甾醇,主要作用于肿瘤抑制蛋白53(TP53)、丝裂原活化蛋白激酶3(MAPK3)等核心靶点。GO富集分析和KEGG通路富集分析结果表明,通塞颗粒治疗AECOPD与TNF、IL-17等信号通路干预氧化应激、减少细胞凋亡、抑制炎症反应有关。分子对接结果显示,通塞颗粒的关键活性成分与治疗AECOPD的核心靶点均有较好的结合活性。动物实验研究结果显示,模型组FVC、FEV_(0.3)、FEV_(0.3)/FVC、PEF值均较Objective:To explore the mechanism of action of Tongsai Granules(TSG)in the treatment of acute exacerbation of chronic obstructive pulmonary disease(AECOPD)based on network pharmacology,molecular docking technology,and animal experiments.Methods:①Predictive analysis.The active ingredients and corresponding targets of TSG were screened by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.AECOPD-related genes were retrieved by GeneCards,OMiM,and other databases.Venn diagrams were drawn to obtain intersecting drug-disease targets;active ingredient of TsG-intersecting target network and a protein-protein interaction network was constructed and core targets were screened out based on network relationships;R programming language software was used for Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Molecular docking of key active ingredients of TSG and core targets Was performed by using AutoDockTool 1.5.6 software.@Experimental verification.Fifty rats were randomly divided into control group,model group,Chinese medicine group,western medicine group,and combination group,with 10 rats in each group.The AECOPD rat model was constructed in all groups except the control group.And received drug gavage 2 days before and 4 days after the acute exacerbation.After treatment,the lung function indicators,including forced expiratory volume in O.3 seconds(FEV_(0.3)),force vital capacity(FVC),the ratio of forced expiratory volume in 0.3 seconds to forced vital capacity(FEV_(0.3)/FVC),and peak expiratory flow(PEF),were measured in each group.The contents of tumor necrosis factor(TNF)-αand interleukin(IL)-7 in serum were measured;the pathological changes in airway and lung tissues were observed by using hematoxylin-eosin(HE)staining and the network pharmacological prediction results were validated by western blotting(WB)and enzyme-linked immunosorbent assay(ELiSA).Results:In the treatment of AECOPD,the key active ingredients of TSG were quer

关 键 词：慢性阻塞性肺疾病急性加重期 通塞颗粒 网络药理学 分子对接技术 核心靶点 信号通路 

分 类 号：R285[医药卫生—中药学]