小儿咳喘颗粒治疗儿童支气管肺炎的活性成分及作用机制预测分析  

Predictive Analysis of the Active Components and Mechanism of Xiaoer Kechuan Granule in the Treatment of Children Bronchopneumonia

作　　者：任素娟 周瑞[1] 唐志书[1] 张海潮 刘妍如[1] 胡锦航 宋忠兴[1] 倪健[2] 石丽 王文宏 Ren Sujuan;Zhou Rui;Tang Zhishu;Zhang Haichao;Liu Yanru;Hu Jinhang;Song Zhongxing;Ni Jian;Shi Li;Wang Wenhong(Shaanxi University of Chinese Medicine/Co-construction Collaborative Innovation Center for Chinese Medicine Resources Industrialization by Shaanxi&Education Ministry/State Key Laboratory of Research&Development of Characteristic Qin Medicine Resources(Cultivation)/Shaanxi Innovative Drug Research Center,Xianyang 712083,China;School of Chinese Materia Medica,Beijing University of Chinese Medicine,Bejing 102488,China;Xi'an Beilin Pharmaceutical Co.,Ltd,Xi'an 712000,China)

机构地区：[1]陕西中医药大学/陕西中药资源产业化省部共建协同创新中心/秦药特色资源研究开发国家重点实验室(培育)/陕西省创新药物研究中心,咸阳712083 [2]北京中医药大学中药学院,北京102488 [3]西安碑林药业股份有限公司,西安712000

出　　处：《世界科学技术-中医药现代化》2023年第1期359-371,共13页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：咸阳市科学技术局2020年度咸阳市重点研发科技专项(2020k02-85):四季抗病毒合剂/消炎退热合剂治疗新型冠状病毒感染肺炎及呼吸道感染疾病的实验研究,负责人:唐志书。

摘　　要：目的利用网络药理学、分子对接、细胞实验探索小儿咳喘颗粒治疗儿童支气管肺炎的活性成分及其作用机制。方法通过TCMSP、TCMID、STITCH数据库和文献检索,收集小儿咳喘颗粒处方中麻黄、川贝母、苦杏仁、黄芩等15味药的活性成分及其作用靶点,并在Genecards和OMIM数据库中查找儿童支气管肺炎靶点。利用STRING网络平台对共有靶点构建蛋白互作网络以及GO(Gene Ontology)功能注释和KEGG(Kyotoencyclopediaofgenesandgenomes)通路分析,并用Cytoscape3.7.2软件构建PPI(Protein-Protein interaction network,PPI network)和“疾病-药物-成分-靶点-通路”网络图,最后根据网络图的分析结果选取关键成分和靶点进行分子对接。体外细胞实验采用脂多糖(Lipopolysaccharide,LPS)诱导的RAW264.7炎症细胞模型,检测小儿咳喘颗粒对RAW264.7细胞的抗炎作用。免疫印迹(Western blot)实验检测肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)、白细胞介素-6(Interleukin 6,IL-6)、白细胞介素-1β(Interleukin-1β,IL-1β)以及血管内皮生长因子A(Vascular endothelial growth factor A,VEGFA)的表达,进一步验证其作用机制。结果收集到小儿咳喘颗粒活性成分有槲皮素、β-谷甾醇、木犀草素、山柰酚等220个,靶点有TNF、IL-6、IL-1β、VEGFA等439个,疾病靶点350个。GO功能和KEGG通路分析表明小儿咳喘颗粒治疗支气管肺炎主要包括IL-17信号通路、AGE-RAGE信号通路与TNF信号通路等,主要涉及细胞代谢、免疫反应和细胞因子转化等生物过程。分子对接结果显示小儿咳喘颗粒的关键成分槲皮素、β-谷甾醇、木犀草素、山柰酚与疾病靶点TNF、IL-6、IL-1β、VEGFA有较强的结合活性。实验结果显示,小儿咳喘颗粒给药组可显著降低炎症因子一氧化氮(NO)、TNF-α、IL-6的含量,同时抑制TNF-α、IL-6、IL-1β以及VEGFA蛋白的表达,初步验证了预测结果的可靠性。结论采用网络药理学结合分子�Objective To explore the active ingredients and their potential mechanism action of Xiaoer Kechuan Granule in the treatment of children bronchopneumonia by the method of network pharmacology,molecular docking and cell experiments.Methods Through TCMSP,TCMID,STITCH database and literature search,to collect the active ingredients and targets of 15 traditional Chinese medicines like Ephedra,Fritillaria cirrhosa D.Don,Semen Armeniacae Amarum,Scutellaria baicalensis Georgi in Xiaoer Kechuan Granule,and collect the disease targets of children bronchopneumonia in Genecards and OMIM database.Through the STRING network platform to build protein interaction network for the common targets,GO(Gene Ontology) functional enrichment and KEGG(Kyoto encyclopedia of genes and genomes) pathway enrichment analysis,and then used the Cytoscape 3.7.2 software to build Protein-Protein interaction network(PPI network) and "disease-drug-ingredients-targets-pathway" network.Finally,according the analysis results of network,make the molecular docking with the key ingredients and targets.In vitro cell test,Lipopolysaccharide(LPS) induced RAW264.7 inflammatory cell model was used to detect the anti-inflammatory effect of Xiaoer Kechuan Granule on RAW264.7 cells,and carried out western blot to detect the expression of Tumor necrosis factor-α(TNF-α),Interleukin 6(IL-6),Interleukin-1β(IL-1β) and Vascular endothelial growth factor A(VEGFA).Results There are 220 different kinds of active ingredients in Xiaoer Kechuan Granule including quercetin,β-sitosterol,luteolin,Kaempferol,etc.and 439 corresponding targets,such as TNF,IL-6,IL-1β and VEGFA.and 350disease targets.GO function and KEGG pathway analysis showed that the treatment of children bronchopneumonia with Xiaoer Kechuan Granule mainly includes IL-17 signaling pathway,AGE-RAGE signaling pathway and TNF signaling pathway,etc.which are mainly involved in biological processes such as cell metabolism,immune response and cytokine conversion.Molecular bonding results showed that quercetin,β-s

关 键 词：小儿咳喘颗粒 儿童支气管肺炎 网络药理学 分子对接 实验验证 

分 类 号：R-058[医药卫生]