基于超高效液相色谱-四级杆-静电场轨道阱串联质谱联用技术结合网络药理学分析仙灵骨葆胶囊的化学成分和作用机制  

Analysis of the chemical components and mechanism of Xianling Gubao Capsules based on UHPLC Q-Exactive Orbitrap-MS combined with network pharmacology

作　　者：程雪怡 刘文 陈梅 陆苑[1] 金阳 李勇军[3] CHENG Xue-yi;LIU Wen;CHEN Mei;LU Yuan;JIN Yang;LI Yong-jun(State Key Laboratory of Functions and Applications of Medicinal Plants,Guizhou Provincial Key Laboratory of Pharmaceutics,Guizhou Medical University,Guiyang 550004,China;School of Pharmacy,Guizhou Medical University,Guiyang 550004,China;Engineering Research Center for the Development and Application of Ethnic Medicine and TCM(Ministry of Education),Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学省部共建药用植物功效与利用国家重点实验室/贵州省药物制剂重点实验室,贵阳550004 [2]贵州医科大学药学院,贵阳550004 [3]贵州医科大学民族药与中药开发应用教育部工程研究中心,贵阳550004

出　　处：《中国新药杂志》2025年第7期747-760,共14页Chinese Journal of New Drugs

基　　金：国家自然科学基金资助项目(82060704)。

摘　　要：目的:通过超高效液相色谱-四级杆-静电场轨道阱串联质谱联用技术结合网络药理学,研究仙灵骨葆胶囊(Xianling Gubao capsules,XLGBC)治疗骨质疏松(osteoporosis,OP)的有效化学成分及潜在作用机制。方法:采用Hypersil Gold色谱柱,以0.1%甲酸水溶液(A)-0.1%甲酸乙腈溶液(B)作为流动相进行梯度洗脱;质谱在正、负离子模式下扫描;建立XLGBC化学成分数据库,结合Xcalibur软件,对比准分子离子和特征碎片离子,进行化学成分鉴定;利用TCMSP和SwissADME数据库筛选候选活性成分,并通过GeneGards,DisGeNET,SwissTargetPrediction和Metascape数据库对活性成分治疗OP的靶点进行预测和富集分析,再结合String平台和Cytoscape软件获取XLGBC抗OP的关键靶点和重要活性成分,利用分子对接技术进行初步验证。结果:共鉴定出120个化学成分,其中5个成分首次在XLGBC中鉴定得到;120个化学组分中筛选到63个治疗OP的候选活性成分和447个靶点,富集分析显示主要涉及激素水平调节、破骨细胞分化、PI3K-AKT信号通路等途径,其中EGFR和PIK3R1靶点与多条通路联系密切,被视为关键靶蛋白;通过蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络拓扑分析获取到XLGBC治疗OP的6个主要核心成分,分子对接表明该成分与关键靶蛋白均具有较强烈的结合活性。结论:全面阐明了XLGBC中的各类化学成分,并揭示了XLGBC作用于OP的主要化学成分和潜在作用机制,为XLGBC的药效物质基础研究提供科学参考。Objective:To investigate the effective chemical components and potential mechanisms of Xianling Gubao Capsules(XLGBC) in the treatment of Osteoporosis(OP),we employed UHPLC Q-Exactive Orbitrap-MS in combination with network pharmacology.Methods:The Hypersil Gold column was used for gradient elution utilizing a mobile phase consisting of 0.1% formic acid in water(A) and a formic acid acetonitrile solution(B).Mass spectrometry was conducted in both positive and negative ion modes.A chemical composition database for XLGBC was established,combining Xcalibur software to compare excimer ion and characteristic fragment ions,thereby chemical components were identified.Candidate active ingredients were screened using TCMSP and SwissADME.The Genegards,DisGeNET,SwissTargetPrediction and Metascape databases were used to predict and enrichg the targets of active ingredients for the treatment of OP.By combining String platform and Cytoscape software,key targets and important active components of XLGBC for anti-OP were obtained.And the molecular docking was used for preliminary verification purposes.Results:A total of 120 chemical components were identified,among which 5 compounds were identified in XLGBC for first time.Additinally,63 candidate active ingredients and 447 targets for the treatment of OP were screened.Enrichment analysis showed that regulation of hormone levels,osteoclast differentiation,PI3K-AKT signaling pathway,etc.,are involved.The EGFR and PIK3R1 targets are closely related to multiple pathways,and were considered as the key target protein.The 6 main core components of XLGBC therapy for OP were obtained via network topology analysis of protein-protein interaction(PPI).And molecular docking indicated that the components have strong binding capacity to the key target proteins.Conclusion:The comprehensive elucidation of various chemical compositions in XLGBC reveals the main chemical components and potential mechanisms of XLGBC on OP,which provides a scientific reference for its effective substances research.

关 键 词：仙灵骨葆胶囊 化学成分 骨质疏松 超高效液相色谱-四级杆-静电场轨道阱串联质谱联用技术 网络药理学 分子对接 

分 类 号：R965[医药卫生—药理学]