摘要
目的 基于超高效液相色谱串联四极杆静电场轨道阱质谱(UPLC-Q Exactive Orbitrap-HRMS)技术、网络药理学和分子对接探究黄连上清丸解热抗炎的活性成分及其作用机制。方法 根据质谱信息、对照品裂解规律及参考文献资料对黄连上清丸中的化学成分进行指认;利用SwissADME数据库筛选黄连上清丸中的成药性成分,PharmMapper数据库获取成药性成分相关作用靶点;利用Gene cards和Disgenet数据库筛选解热、炎症相关靶点;对得到的成分作用靶点与疾病相关靶点取交集,获得黄连上清丸发挥解热抗炎作用的潜在靶点,采用Cytoscape 3.10.1软件构建“成分-靶点-疾病-复方”网络,分析活性成分;将共有靶点导入STRING网站,构建蛋白质-蛋白质相互作用(PPI)网络,分析关键靶点;通过微生信网站对关键靶点进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析;将活性成分与关键靶点进行分子对接验证;通过建立HPLC指纹图谱,控制黄连上清丸的质量。结果 从黄连上清丸中共鉴定144个化合物,其中黄酮类58个、生物碱类17个、环烯醚萜类7个、木脂素类4个、色原酮类8个、蒽醌类7个、香豆素类2个、皂苷类5个、有机酸类5个、酚酸类7个、苯丙素类9个、苯乙醇苷类4个、糖类2个、内酯类1个,以及其他类8个。黄连上清丸发挥解热抗炎作用的潜在作用靶点有218个,涉及2 036个生物过程、119个细胞组分、281个分子功能和155条信号通路。筛选得到莲心季铵碱、甘草宁G、大黄素、氧化小檗碱、甘草素、大黄素甲醚、花椒毒酚、粘毛黄芩素Ⅲ等12个潜在活性成分,胰岛素(INS)、RAC-α丝氨酸/苏氨酸蛋白激酶(AKT1)、表皮生长因子受体(EGFR)、热休克蛋白HSP 90-α(HSP90AA1)、基质金属蛋白酶-9(MMP9)等10个关键靶蛋白,以及Ras信号通路、叉头框蛋白O信号通路、Rap1信号通路、丝裂原活化蛋白激酶信号通路等信号通路。12个潜在活性成分与关键靶点有较强的结合活性。建立的指纹图谱可指认黄连上清丸中的8个活性成分。结论 黄连上清丸可能是通过莲心季铵碱、甘草宁G、大黄素、氧化小檗碱、甘草素、大黄素甲醚等活性成分,作用于INS、AKT1、EGFR、HSP90AA1、MMP9等关键靶点,通过Ras信号通路、叉头框蛋白O信号通路、Rap1信号通路、丝裂原活化蛋白激酶信号通路等信号通路,发挥解热抗炎作用。
Objective To investigate the active components and its mechanism for the effect on antipyretic and anti-inflammatory in Huanglian Shangqing Pills using UPLC-Q Exactive Orbitrap-HRMS,network pharmacology and molecular docking.Methods According to the mass spectrometry information,the cleavage law of the reference substance and the reference literatures,the chemical compositions in Huanglian Shangqin Pills were identified;SwisSADME database was used to screen the druggable components in Huanglian Shangqing Pills,and the PharmMapper database was used to obtain the relevant targets of the druggable components;Antipyretic and inflammationrelated targets was screened by Gene cards and Disgenet databases;The intersections between the obtained component targets and diseaserelated targets were used to obtain the potential targets of Huanglian Shangqing Pills to exert antipyretic and anti-inflammatory effects,and the"component-target-disease-compound"network was constructed by Cytoscape 3.10.1 software to analyze the active ingredients;The common targets were imported into the String website for protein-protein interaction analysis to search for key targets;GO function and KEGG pathway enrichment analysis of key targets were carried out through bioinformatics website;Molecular docking was used to validate the active ingredients and key targets;The quality of Huanglian Shangqing Pills was controlled by establishing HPLC fingerprints.Results A total of 144 compounds were identified in Huanglian Shangqing Pills,including 58 flavonoids,17 alkaloids,7 iridoid terpenoids,4 lignans,8 chromogenone,7 anthraquinones,2 coumarins,5 saponins,5 organic acids,7 phenolic acids,9 phenylpropanoids,4 phenylethanol glycosides,2 sugars,1 lacone,and 8 other substances.There were 218 potential targets for antipyretic and anti-inflammatory effects,involving 2036 biological processes,119 cellular components,281 molecular functions and 155 signaling pathways.Twelve potential active ingredients including lotusine,gancaonin G,emodin,berlambine,glycyrrhizin,physcion,xanthotoxol,ganhuangenin and others,10 key target proteins such as INS,AKT1,EGFR,HSP90AA1,MMP9 and so on,and signaling pathways such as Ras signaling pathway,FoxO signaling pathway,Rap1 signaling pathway,and MAPK signaling pathway were screened.The 12 potential active ingredients have strong binding activity to key targets.The established fingerprint identified eight active ingredients in Huanglian Shangqing Pills.Conclusion Huanglian Shangqing Pills may act on key targets such as INS,AKT1,EGFR,HSP90AA1,MMP9,etc.through active components such as lotusine,gancaonin G,emodin,berlambine,glycyrin,physcion,etc.,and play the role through the signaling pathways such as Ras signaling pathway,FoxO signaling pathway,Rap1 signaling pathway,and MAPK signaling pathway,etc.
作者
曲婷丽
王二兵
张楠
许信学
周小雪
高耀
QU Tingli;WANG Erbing;ZHANG Nan;XU Xinxue;ZHOU Xiaoxue;GAO Yao(School of Pharmaceutical Science,Shanxi Medical University,Taiyuan 030001,China;College of Chemical Engineering and Technology,Taiyuan University of Science and Technology,Taiyuan 030024,China;Shanxi Huakang Pharmaceutical Co.,Ltd.,Yuncheng 044200,China;Department of Psychiatry,First Hospital/First Clinical Medical College of Shanxi Medical University,Taiyuan 030001,China)
出处
《药物评价研究》
北大核心
2025年第2期394-415,共22页
Drug Evaluation Research
基金
国家自然科学基金青年项目(82301725)
中国博士后科学基金第73批面上资助项目(2023M732155)
山西省科技厅基础研究项目(20210302124586)
山西省重点研发项目(202102130501023)
太原市科技局“双百攻关行动”首批关键核心技术攻关“揭榜挂帅”项目(2024TYJB0147)
山西省技术创新中心项目(202104010911006)
山西省教育厅教改项目(J20230518)。
