基于液质联用结合网络药理学的复方伤痛胶囊中5种药效成分定量及作用机制研究  

Quantitative analysis and mechanism research of five pharmacodynamic components in Fufang Shangtong Capsule based on liquid mass spectrometry combined with network pharmacology

作　　者：高渐联[1] 周霖[2] 王肖辉 聂兴国[3] 朱超男 GAO Jian-lian;ZHOU Lin;WANG Xiao-hui;NIE Xing-guo;ZHU Chao-nan(Department of Pharmacy,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,China;Department of Pharmacy,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Orthopedics,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,China)

机构地区：[1]新乡医学院第一附属医院药学部,河南卫辉453100 [2]郑州大学第一附属医院药学部,河南郑州450052 [3]新乡医学院第一附属医院骨科,河南卫辉453100

出　　处：《中草药》2023年第6期1793-1803,共11页Chinese Traditional and Herbal Drugs

基　　金：北京医卫健康公益基金会(TM19004);河南省医学攻关计划(LHGJ20200524);河南省科技攻关计划(21210231107)。

摘　　要：目的对复方伤痛胶囊中的5种药效成分进行定量,提升药物质量标准,并对药效成分在药物治疗中的作用机制进行研究。方法采用Acquity UPLC^(■)BEH C_(18)色谱柱(50 mm×2.1 mm,1.7μm)进行成分初步分离,乙腈-0.1%甲酸水溶液作为流动相梯度洗脱;采用UHPLC-Q-Orbitrap HRMS仪器,使用Full mass/dd MS2模式对药物中的关键成分进行快速、准确含量测定;基于网络药理学构建关键成分-重要靶点网络,并进一步进行京都基因和基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)和基因本体(gene ontology,GO)分析,找到关键信号通路和作用靶点。结果建立的分析方法学较为稳定,能够满足分析的标准需要;5种药效成分(没食子酸、大黄素、大黄酸、大黄素甲醚和延胡索乙素)线性关系均良好,r≥0.9990;网络药理学结果显示,药物中的药效成分可能是通过基质金属蛋白酶9(matrix metallo protein 9,MMP9)、前列腺素内过氧化物合酶2(prostaglandin endoperoxide synthase 2,PTGS2)、MMP2、白细胞介素-2(interleukin-2,IL-2)、表皮生长因子受体(epidermal growth factor receptor,EGFR)等炎症、疼痛关键靶点,从而发挥缓解疼痛及炎症等作用。实验验证结果发现,没食子酸能够明显降低脂多糖诱导的小鼠单核巨噬细胞白血病RAW264.7细胞中的IL-2水平,大黄素能够明显降低RAW264.7细胞中的PTGS2水平,大黄酸能够明显降低RAW264.7细胞中的MMP9水平,大黄素甲醚能够明显降低RAW264.7细胞中的EGFR水平,延胡索乙素能够明显降低RAW264.7细胞中的MMP2水平。结论对复方伤痛胶囊的质量提升提供了重要的参考依据,同时为中医药相关药效成分研究提供了一种借鉴思路。Objective To accurately quantify the five pharmacodynamic components in Fufang Shangtong Capsule(复方伤痛胶囊,FSC),improve drug quality standards,and analyze the important role of pharmacodynamic components in drug therapy.Methods An Acquity UPLC^(■)BEH C_(18)column(50 mm×2.1 mm,1.7μm)was used to separate the components.Acetonitrile-0.1%formic acid water was used as mobile phase gradient elution.UHPLC-Q-Orbitrap HRMS and Full Mass/dd MS2 model were used for rapid and accurate content determination of key ingredients in the drug.Key components-important target network was constructed based on network pharmacology,and Kyoto encyclopedia of genes and genomes(KEGG)and gene ontology(GO)analysis was further conducted to find key signaling pathways and action targets.Results The analytical methodology established in this study was robust and could meet the standard requirements of analysis.All the five pharmacodynamic components had good linearity in the range of mass concentration,r≥0.9990.Network pharmacology results showed that the pharmacodynamic components in drugs may be key targets of inflammation and vascular factors such as matrix metallo protein 9(MMP9),prostaglandin endoperoxide synthase 2(PTGS2),MMP2,interleukin-2(IL-2),epidermal growth factor receptor(EGFR).As a result,it maps to nuclear factor-κB(NF-κB),transforming growth factor-β(TGF-β),tumor necrosis factor(TNF)signaling pathway inflammatory pathways,which has important reference value in drug quality and efficacy.The experimental results showed that gallic acid could significantly reduce the level of IL-2 in LPS induced RAW264.7 cells,emodin could significantly reduce the level of PTGS2 in cells,rhein could significantly reduce the level of MMP9 in cells,emodin methyl ether could significantly reduce the level of EGFR in cells,and fumarine ethyl rhizoma could significantly reduce the level of MMP2 in cells.Conclusion This study provides an important reference for the quality improvement of FSC,and also provides a reference for the study

关 键 词：网络药理学 复方伤痛胶囊 没食子酸 大黄素 大黄酸 大黄素甲醚 延胡索乙素 基质金属蛋白酶9 前列腺素内过氧化物合酶2 白细胞介素-2 表皮生长因子受体 核因子-κB 转化生长因子-β 肿瘤坏死因子 

分 类 号：R283.6[医药卫生—中药学]