摘要
目的运用UHPLC-QTOF-MS和网络药理学-分子对接技术,探讨青阳参总苷治疗社会挫败的物质基础和潜在的作用机制。方法首先,采用UHPLC-QTOF-MS鉴定青阳参总苷化学成分。其次,运用Pubchem下载青阳参总苷化学成分2D结构后,利用Swiss Target Prediction数据库预测其靶点;从TTD、GeneCards、CTD数据库获取社会挫败相关治疗靶点;通过Venny平台获取两者的交集靶点;采用Cytoscape构建“药物-活性成分-靶点-疾病”网络;运用String数据库构建蛋白互作(PPI)网络后,结合Cytoscape分析得交集靶点排名;使用DAVID数据库对交集靶点进行基因本体(GO)功能注释和京都基因与基因组百科全书(KEGG)通路富集分析。利用AutoDock vina软件进行分子对接验证。最后,构建社会挫败小鼠模型,采用ELISA法检测小鼠血清5-羟色胺(5-HT)、多巴胺(DA)含量,HE染色观察小鼠海马组织神经细胞病理变化。结果从青阳参总苷共鉴定152个化学成分。预测及筛选得到丝氨酸/苏氨酸蛋白激酶(AKT1)、单胺氧化酶A(MAOA)、多巴胺受体D3(DRD3)等26个青阳参总苷治疗社会挫败潜在作用靶点,对应85个青阳参总苷活性成分;富集分析得到45个GO注释条目(P<0.01)、11条KEGG信号通路。分子对接发现青阳参苷元、白桦脂酸、绿原酸等20个活性成分与AKT1、DRD4、DRD2、DRD3等10个核心靶蛋白均可自发结合。动物实验表明,与正常对照组相比,模型对照组小鼠血清中5-HT、DA含量均明显降低(P<0.05);小鼠海马组织神经细胞排列疏松,有明显的细胞丢失。与模型对照组相比,青阳参总苷各剂量组小鼠血清中5-HT、DA含量均显著升高(P<0.05,P<0.01);青阳参总苷中、高剂量组小鼠海马组织神经细胞间隙减小,且排列较为整齐。结论青阳参总苷对社会挫败具有治疗作用。主要药效物质为青阳参苷元、白桦脂酸、绿原酸等,可通过多靶点、多通路发挥抗社会挫败作用。
Objective The present study explored the material basis and mechanism of Qingyangshen glycosides in treatment of social defeat based on UHPLC-QTOF-MS and network pharmacology-molecular docking techniques.Methods Firstly,UHPLC-QTOF-MS was used to identify the chemical constituents of Qingyangshen glycosides.Secondly,after the 2D structure of the chemical components of Qingyangshen glycosides was downloaded by Pubchem,the chemical target of Qingyangshen glycosides was predicted by swiss target Prediction database;TTD,genecards and CTD databases were used to obtain therapeutic targets related to social defeat;he intersection targets of the two were obtained through Venny platform;Cytoscape was used to construct a"drug-active component-target-disease"network;After protein interaction network(PPI)was constructed by using String database,the intersection targets were ranked by Cytoscape analysis;gene ontology(GO)functional annotation and genome encyclopedia(KEGG)pathway enrichment analysis were performed for intersection targets using DAVID database;AutoDock Vina software was used for molecular docking verification.Finally,a mouse model of social defeat was established,the levels of 5-hydroxytryptamine(5-HT)and dopamine(DA)in serum were detected by ELISA,and the pathological changes of neurons in the hippocampus were observed by HE staining.Results A total of 152 chemical components were identified from Qingyangshen glycosides.After prediction and screening,26 potential anti-social defeat targets including serine/threonine protein kinase(AKT1),monoamine oxidase A(MAOA)and dopamine D3 receptor(DRD3)were obtained,corresponding to 85 active components of Qingyangshen glycosides;45 GO annotation items and 11 KEGG signaling pathways were obtained by enrichment analysis of anti-social frustration intersection targets of Qingyangshen glycosides.Molecular docking showed that Qingyangshengenin,betulinic acid,chlorogenic acid,could stably bind to AKT1,DRD4,DRD2 and DRD3.Compared with normal control group,the contents of 5-HT and DA in serum of model control group were significantly decreased(P<0.05);The arrangement of nerve cells in the hippocampus of mice was loose and obvious cell loss was observed.Compared with model control group,the contents of 5-HT and DA in serum of fluoxetine positive control group and qingyangshen glycosides low-dose,medium-dose and high-dose groups were significantly increased(P<0.05,P<0.01);the intercellular Spaces of hippocampal tissues in medium and high dose groups of Qingyangshen glycosides were decreased and the arrangement was more orderly.Conclusion Qingyangshen glycosides has therapeutic effect on social defeat.The main pharmacoactive substances are qingyangshenogenin,betulinic acid,chlorogenic acid,etc.,which can exert anti-social defeat effect through multi-target and multi-pathway.
作者
王静茹
刘顶鼎
常露露
朱蔷
杨菁
吴雪梅
曾贵荣
Wang Jingru;Liu Dingding;Chang Lulu;Zhu Qiang;Yang Jing;Wu Xuemei;Zeng Guirong(Guizhou University of Traditional Chinese Medicine,School of Pharmacy&Research Center for Pharmacodynamic Material Basis and Mechanism of Action,Guiyang 550025,China;the First Affiliated Hospital of Guizhou University of TCM,Guiyang 550001,China;Hunan Key Laboratory of Pharmacodynamics and Safety Evaluation of New Drugs&Hunan Provincial Research Center for Safety Evaluation of Drugs,Changsha 410331,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第2期564-579,共16页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
贵州中医药大学国家自然科学基金后补助资金科研创新探索专项项目(2018YFC170810502):基于BDNF-TrkB/CREB/ERK信号通路探讨青阳参提取物对社会挫败模型小鼠的作用及机制研究,负责人:刘顶鼎
贵州省卫生健康委员会科学技术基金项目(gzwkj2023-147):基于PGC1α/FNDC5/BDNF通路调节神经可塑性探讨C21甾体皂苷抗社会挫败的作用机制,负责人:刘顶鼎
贵州省中医药管理局中医药、民族医药科学技术研究项目(QZYY-2022-008):青阳参提取物对社会挫败小鼠海马组织PGC1α/FNDC5/BDNF信号通路和突触可塑性的影响,负责人:刘顶鼎
作者简介
通讯作者:刘顶鼎,副教授,主要研究方向:中药、民族药及复方防治神经精神系统疾病;通讯作者:曾贵荣,副研究员,主要研究方向:神经精神药理。
