UPLC-Q-Exactive-MS/MS联合网络药理学初探金乌骨通胶囊治疗骨质疏松症的活性成分及作用机制  

Active components and mechanism of Jinwugutong Capsules in treatment of osteoporosis:a study based on UPLC-Q-Exactive-MS/MS combined with network pharmacology

作　　者：杨小双 宋信莉 宋学丽 杨应勇 刘文 杨胜磊 石佳楠 沈丽 万开龙 刘兴德 YANG Xiao-shuang;SONG Xin-li;SONG Xue-li;YANG Ying-yong;LIU Wen;YANG Sheng-lei;SHI Jia-nan;SHEN Li;WAN Kai-long;LIU Xing-de(School of Pharmacy,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;National Engineering Research Center of Miao's Medicines,Guiyang 550025,China;Guizhou Chinese Medicine Processing and Preparation Engineering Technology Research Center,Guiyang 550025,China;Guizhou Shenqi Pharmaceutical Co.,Ltd.,Guiyang 550004,China;School of Pharmacy,Guizhou Medical University,Guiyang 550025,China)

机构地区：[1]贵州中医药大学药学院,贵州贵阳550025 [2]国家苗药工程技术研究中心,贵州贵阳550025 [3]贵州中药炮制与制剂工程技术研究中心,贵州贵阳550025 [4]贵州神奇药业有限公司,贵州贵阳550004 [5]贵州医科大学药学院,贵州贵阳550025

出　　处：《中国中药杂志》2023年第12期3360-3372,共13页China Journal of Chinese Materia Medica

基　　金：贵州省特色功能食品与中药制剂开发攻关大平台黔教合项目(KY字[2020]006);贵州中医药大学药用高分子材料研究中心项目(贵中医党办发[2019]70号);贵州省国内一流学科建设项目(GNYL[2017]008);贵州省药物新剂型新工艺科技创新人才团队项目(黔科合平台人才[2017]5655)。

摘　　要：基于UPLC-Q-Exactive-MS/MS技术,联合网络药理学初步探讨金乌骨通胶囊治疗骨质疏松症的活性成分及作用机制。首先采用UPLC-Q-Exactive-MS/MS对金乌骨通胶囊化学成分进行表征,并通过网络药理学的研究方法构建“药物-成分-靶点-通路-疾病”网络,获取关键靶点及主要活性成分。其次,应用AutoDock软件对主要活性成分与关键靶点进行分子对接验证。最后,建立骨质疏松动物模型,采用酶联免疫吸附法(ELISA)测定金乌骨通胶囊对关键靶点AKT1、ALB、TNF-α表达的影响。从金乌骨通胶囊中共鉴定出化学成分59个,其中补骨脂甲素、8-异戊烯基柚皮素、去甲氧基姜黄素、异补骨脂二氢黄酮、金雀异黄酮等可能为金乌骨通胶囊治疗骨质疏松症的主要活性成分。蛋白-蛋白互作(PPI)网络拓扑分析得到AKT1、ALB、CTNNB1、TNF、EGFR等10个核心靶点。KEGG富集显示金乌骨通胶囊主要通过调控PI3K-AKT信号通路、神经活性配体-受体相互作用、MAPK信号通路、Rap1信号通路等发挥作用。分子对接显示金乌骨通胶囊主要活性成分与关键靶点结合良好,ELISA结果表明金乌骨通胶囊可以下调AKT1、TNF-α水平,上调ALB水平,初步验证了网络药理学的可靠性。该研究表明金乌骨通胶囊可能通过“多成分-多靶点-多途径”发挥治疗骨质疏松症的作用,可为后续深入研究提供参考。UPLC-Q-Exactive-MS/MS and network pharmacology were employed to preliminarily study the active components and mechanism of Jinwugutong Capsules in the treatment of osteoporosis.Firstly,UPLC-Q-Exactive-MS/MS was employed to characterize the chemical components of Jinwugutong Capsules,and network pharmacology was employed to establish the"drug-component-targetpathway-disease"network.The key targets and main active components were thus obtained.Secondly,AutoDock was used for the molecular docking between the main active components and key targets.Finally,the animal model of osteoporosis was established,and the effect of Jinwugutong Capsules on the expression of key targets including RAC-alpha serine/threonine-protein kinase(AKT1),albumin(ALB),and tumor necrosis factor-alpha(TNF-α)was determined by enzyme-linked immunosorbent assay(ELISA).A total of 59 chemical components were identified from Jinwugutong Capsules,among which coryfolin,8-prenylnaringenin,demethoxycurcumin,isobavachin,and genistein may be the main active components of Jinwugutong Capsules in treating osteoporosis.The topological analysis of the protein-protein interaction(PPI)network revealed 10 core targets such as AKT1,ALB,catenin beta 1(CTNNB1),TNF,and epidermal growth factor receptor(EGFR).The Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment showed that Jinwugutong Capsules mainly exerted the therapeutic effect by regulating the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling pathway,neuroactive ligand-receptor interaction,mitogen-activated protein kinase(MAPK)signaling pathway,Rapl signaling pathway and so on.Molecular docking showed that the main active components of Jinwugutong Capsules well bound to the key targets.ELISA results showed that Jinwugutong Capsules down-regulated the protein levels of AKT1 and TNF-αand up-regulated the protein level of ALB,which preliminarily verified the reliability of network pharmacology.This study indicates that Jinwugutong Capsules may play a role in the treatment of osteoporosis th

关 键 词：金乌骨通胶囊 骨质疏松症 UPLC-Q-Exactive-MS/MS 网络药理学 分子对接 

分 类 号：R285[医药卫生—中药学]