基于网络药理学和体内实验研究五虎汤干预RSV诱导哮喘的效应物质及作用机制  

Active components and mechanism of Wuhu Decoction in intervening RSV-induced asthma based on network pharmacology and in vivo experiment

作　　者：姚冰 刘靖雷 王孟清[2] 罗银河[1] 谢静[2] 荀春铮 李凡[2] 黄婷[2] 曾庆佳[2] YAO Bing;LIU Jing-lei;WANG Meng-qing;LUO Yin-he;XIE Jing;XUN Chun-zheng;LI Fan;HUANG Ting;ZENG Qing-jia(Hunan University of Traditional Chinese Medicine,Changsha 410208,China;the First Afiliated Hospital of Hunan University of Traditional Chinese Medicine,Changsha 410021,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南中医药大学第一附属医院,湖南长沙410021

出　　处：《中国中药杂志》2024年第15期4148-4157,共10页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82174437);全国名老中医药专家传承工作室项目(国中医药人教函[2022]75号);湖南省“十四五”第二批中医药领军人才项目(湘中药发[2023]4号);湖南省自然科学基金项目(2022JJ30037);湖南中医药大学中医儿科学湖南省“双一流”学科建设项目(教高厅函[2022]14号);湖南中医药大学校级科研基金项目(2022CX27);湖南省中医药管理局重点课题(A2024007)。

摘　　要：探讨五虎汤治疗呼吸道合胞病毒(RSV)诱发哮喘的效应物质及作用机制。使用超高效液相串联高分辨质谱仪确定五虎汤的入血成分;借助数据库对入血成分作用于哮喘的靶点进行京都基因与基因组百科全书(KEGG)富集分析及基因本体论(GO)功能分析。同时将靶点蛋白及代谢物-靶点-通路等信息导入STRING数据库,构建蛋白互作网络,明确五虎汤核心成分及关键通路。体内实验部分使用RSV联合鸡卵清蛋白(OVA)建立哮喘模型,通过肺功能、苏木精-伊红(HE)染色、酶联免疫吸附测定(ELISA)法、蛋白免疫印迹法及免疫组织化学法验证五虎汤对RSV诱导哮喘小鼠的干预作用。结果显示,五虎汤入血成分以黄酮类、苯丙素类、木脂素类、萜类等化合物为主,作用于儿童哮喘的核心成分有(-)-表没食子儿茶素、山柰素、异甘草素、香叶木素、桦木酸、熊果酸、瑞香素、七叶素等,钙信号通路、神经活性配体-受体相互作用、NOD样受体信号通路、T细胞受体信号通路、Toll样受体信号通路等是其靶点主要作用通路。体内实验结果表明,五虎汤能够改善肺功能指标,下调白细胞介素-6(IL-6)、白细胞介素-17((IL-17)及肿瘤坏死因子-α(TNF-α)水平,并且能够降低肺组织中NOD样受体热蛋白结构域相关蛋白3(NLRP3)、丝裂原活化蛋白激酶1(MAPK1)、丝裂原活化蛋白激酶14(MAPK14)、核因子-κB亚基1(NFKB1)等蛋白的表达,减轻中性粒细胞炎症浸润及肺淤血。结果表明,五虎汤通过多组分、多靶点、多途径的协同作用干预病毒诱发哮喘,体内实验证明其能抑制NOD样受体信号通路的激活,减轻哮喘小鼠气道炎症与气道损伤。This study aims to explore the active components and mechanism of Wuhu Decoction in treating respiratory syncytial virus(RSV)-induced asthma.Ultra-high performance liquid chromatography coupled with high-resolution mass spectrometry was used to determine the components of Wuhu Decoction in the blood.By utilizing databases,Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis and Gene Ontology(GO)functional analysis were conducted to investigate the targets of the components of Wuhu Decoction in asthma.Furthermore,the information on target proteins,and metabolite-target-pathway was imported into the STRING database to construct a network interaction diagram to identify the core components and key pathways of Wuhu Decoction.In the in vivo experiment,an asthma model was established using RSV combined with ovalbumin(OVA)in mice.The intervention effect of Wuhu Decoction on RSV-induced asthma in mice was validated through lung function tests,hematoxylin-eosin(HE)staining,enzyme-linked immunosorbent assay(ELISA),Western blot,and immunohistochemistry.The results showed that the main components of Wuhu Decoction in the blood were flavonoids,phenylpropanoids,lignans,and terpenoids.The core components of Wuhu Decoction in treating pediatric asthma included(-)-epigallocatechin,kaempferol,isoliquiritigenin,diosmetin,betulinic acid,ursolic acid,daphnetin,aescin.The main pathways targeted by Wuhu Decoction were calcium signaling pathway,neuroactive ligand-receptor interaction,NOD-like receptor signaling pathway,T cell receptor signaling pathway,and Toll-like receptor signaling pathway.The results of in vivo experiments demonstrated that Wuhu Decoction could improve lung function indicators,down-regulate levels of interleukin-6(IL-6),interleukin-17(IL-17),and tumor necrosis factor-alpha(TNF-α),and reduce the expression of proteins such as NOD-like receptor pyrin domain-containing 3(NLRP3),mitogen-activated protein kinase 1(MAPK1),mitogen-activated protein kinase 14(MAPK14),and nuclear factor kappaB subunit 1(NFKB1)in

关 键 词：网络药理学 五虎汤 中药入血成分 RSV 儿童哮喘 NOD样受体信号通路 

分 类 号：R285.5[医药卫生—中药学]