目的:探讨小儿定喘颗粒治疗支气管哮喘的药效物质基础与潜在作用机制.方法:采用超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOFMS)对小儿定喘颗粒中的化学成分及入血成分进行分析,结合PubChem、TCMSP、SwissTargetPre-diction数据库获取入血成分的作用靶点;采用GeneCards、OMIM、TTD数据库获取哮喘的靶点信息;应用String平台构建靶蛋白相互作用网络;对交集靶点进行GO、KEGG通路富集分析,利用Auto Dock Vina软件对关键靶点及核心成分进行分子对接验证;利用Gromacs软件对最佳结合模型进行分子动力学模拟实验,采用Western blot法验证小儿定喘颗粒对哮喘富集关键通路相关蛋白表达的影响.结果:共识别出小儿定喘颗粒中107种化学成分(主要包括黄酮类及生物碱类)及小鼠血清中25种原型成分,疾病与入血成分具有269个交集靶点,主要涉及PI3K-Akt、MAPK、EGFR酪氨酸激酶抑制等信号通路,分子对接结果显示核心成分与关键靶点具有较好的结合能,分子动力学模拟结果表明黄芩素与TNF结合稳定且紧密,动物实验结果表明小儿定喘颗粒可以显著抑制哮喘引起的PI3K-Akt信号通路的激活,下调相关蛋白的表达,与网络药理学预测结果基本一致.结论:小儿定喘颗粒中的汉黄芩素、异甘草素、黄芩素、5,6-二羟基-7-甲氧基黄酮及去甲汉黄芩素等成分可通过调节AKT1、TNF、MAPK3等关键靶点干预PI3K-Akt、MAPK、EGFR酪氨酸激酶抑制等信号通路,从而通过多成分、多靶点、多通路的综合作用以有效治疗哮喘.
Exploring potential pharmacodynamic substances basis and mechanism of Xiaoer Dingchuan Granules in the treatment of bronchial asthma based upon UPLC-Q-TOFMS and network pharmacology
OBJECTIVE To explore the pharmacodynamic basis and potential mechanism of Xiaoer Dingchuan Granules(XDG)in the treatment of bronchial asthma(BA).METHODS UPLC-Q-TOFMS was utilized for analyzing the chemical and blood components of XDG and the targets of blood components were obtained by searching the databases of PubChem,TCMSP and SwissTargetPrediction.The databases of GeneCards,OMIM and TTD were employed for acquiring target information for BA;String platform for constructing the target protein interaction network;GO and KEGG pathway enrichment analysis for inter-section targets and key targets and core components verified by molecular docking verification by Auto Dock Vina software.Gro-macs software was utilized for performing molecular dynamic simulation experiments on optimal binding model.And Western blot was used for verifying the effect of XDG on the expression of asthma enrichment key pathway-related proteins.RESULTS A total of 107 chemical components(mostly flavonoids and alkaloids)and 25 prototypic components in murine sera.There were 269 intersection targets between disease and blood components,mainly involving PI3K-Akt,MAPK,EGFR tyrosine kinase inhibi-tion and other signaling pathways.Molecular docking results indicated that core components had excellent binding energy to key targets.CONCLUSION The components of baicalin,isoliquiritigenin,baicalin,5,6-dihydroxy-7-methoxyflavonoid and demethyl baicalin in Xiaoer Dingchuan Granules(XDG)may activate PI3K-Akt/MAPK/EGFR tyrosine kinase inhibition and other signaling pathways through AKT1,TNF,MAPK3 and other key targets to effectively treat BA through the comprehensive effects of multiple components,targets and pathways.
NETL
NSTL
万方数据
