基于UHPLC-Q-TOF-MS整合网络药理学探讨青娥丸效应成分及抗抑郁作用机制  

Study on effective constituents of Qing’e pill and its mechanism of antidepressant activity based on UHPLC-Q-TOF-MS intergrated network pharmacology

作　　者：马程遥 宋珊珊 王一清[1] 赵权 戴国梁[1] 许美娟[1] 居文政[1] MA Cheng-yao;SONG Shan-shan;WANG Yi-qing;ZHAO Quan;DAI Guo-liang;XU Mei-juan;JU Wen-zheng(Dept of Clinical Pharmacology,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;Postdoctoral Fluxion Station,Nanjing University of Chinese Medicine,Nanjing 210023,China;Dept of Pharmacy,Changzhou Hospital Affiliated to Nanjing University of Chinese Medicine,Changzhou,Jiangsu 213003,China)

机构地区：[1]南京中医药大学附属医院临床药理科,江苏南京210029 [2]南京中医药大学博士后流动站,江苏南京210023 [3]南京中医药大学常州附属医院药学部,江苏常州213003

出　　处：《中国药理学通报》2023年第11期2170-2176,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82104355);中国博士后科学基金资助项目(No 2021M701766);江苏省博士后科研资助项目(No 2021K315C)。

摘　　要：目的UHPLC-Q-TOF-MS技术明确青娥丸的入血成分,结合网络药理学探讨青娥丸抗抑郁的有效成分及作用机制。方法采用UHPLC-Q-TOF-MS识别青娥丸入血成分,结合PharmMapper、DisGeNet、GeneCards等数据库以及Cytoscape、ClusterProfiler程序,筛选了青娥丸抗抑郁的核心药效成分与重要靶点,并进行GO及KEGG分析。通过Circlize构建成分-靶点-通路图筛选出核心靶点,应用Autodock vina分子对接结合蛋白免疫印迹法验证。结果青娥丸水提物中的18个入血成分,可干预50个潜在抗抑郁靶点;其中入血成分中9个为可干预多个靶点的核心成分,26个为受多个成分调控的重要靶点;京尼平苷酸、异补骨脂素等9个核心成分可能通过调控26个重要靶点,进而通过PI3K-AKT、雌激素等信号通路发挥抗抑郁作用;EGFR、AKT1为参与多个通路的核心靶点,ESR1和ESR2为受多个核心成分干预的关键靶点;分子对接结果提示,9个核心成分中7个与靶点亲和作用良好;蛋白免疫印迹法证实青娥丸对ESR1和ESR2编码蛋白确有调控作用。结论初步阐明青娥丸抗抑郁的潜在药效物质和作用机制,为筛选青娥丸的抗抑郁药效成分及深入阐明其作用机制提供了科学的理论依据。Aim To elucidate the effective constituents of Qing’e pill and its mechanism of antidepressant activity.Methods UHPLC-Q-TOF-MS technology was employed to identify the constituents migrating to plasma of Qing’e pill.Combined with PharmMapper,DisGeNet,GeneCards databases as well as Cytoscape software,the important active constituents and targets of Qing'e pill were screened.Then,GO and KEGG analysis were used to construct pathway by ClusterProfiler.The core targets were screened by constructing a constituent-target-pathway network by Circlize package.Lastly,Autodock vina was used for molecular docking verification,and Western blotting was employed to verify the regulation of Qing’e pill on the core targets.Results A total of 18 constituents migrating to blood from aqueous extract of Qing’e pill and 50 potential antidepressant targets were obtained from UHPLC-Q-TOF-MS intergrated network pharmacology analysis.Topological analysis suggested that nine constituents were core components which interfered with multiple targets,and twenty-six targets were important targets affected by multiple constituents.Geniposidic acid,isopsoralen and other core constituents may affect 26 important targets,then exerting antidepressant activity through PI3K-AKT,estrogen and other signaling pathways.EGFR and AKT1 were core targets involved in multiple pathways,ESR1 and ESR2 were key targets affected by multiple core components.Molecular docking showed 7 of the 9 core components had a certain degree of affinity with EGFR,AKT1,ESR1 and ESR2,and Western blotting confirmed the regulatory effect of Qing’e pill on proteins encoded by ESR1 and ESR2.Conclusions This study preliminarily clarifies the potential pharmacodynamic constituents and mechanism of Qing’e pill for its antidepressant activity,provides a scientific theoretical basis for screening the antidepressant active constituents of Qing’e pill,and further elucidates its mechanism.

关 键 词：网络药理学 LC-Q-TOF/MS 青娥丸 抑郁症 有效成分 雌激素受体 

分 类 号：R289.5[医药卫生—方剂学] R319[医药卫生—中药学] R392.11[医药卫生—中医学] R749.42