Objective:Gegen Qinlian decoction(GQD)is a classical traditional Chinese medicine formulation which has been used for almost 2000 years.At Guang'anmen Hospital,Beijing,a modified GQD version(m GQD)with seven inste...Objective:Gegen Qinlian decoction(GQD)is a classical traditional Chinese medicine formulation which has been used for almost 2000 years.At Guang'anmen Hospital,Beijing,a modified GQD version(m GQD)with seven instead of four herbal ingredients has been applied to treat Type 2 diabetes.Quality control is a crucial prerequisite for the therapeutic application of herbal medicines.For the identification of products derived from classical GQD,the Chinese Pharmacopeia requires the analysis of only three marker compounds.Because m GQD is a more complex mixture containing seven herbs and hundreds of constituents,the pharmacopoeia method for GQD is inadequate.Materials and Methods:A more comprehensive characterization of the formula's constituents has been developed using ultra-high-performance liquid chromatography-diode array detection(UHPLC-DAD)-Q-Exactive-mass spectrometry(MS)in electrospray ionization positive and negative mode.Moreover,a new method for the fingerprint analysis of m GQD via high-performance thin-layer chromatography(HPTLC)has been established.Results:Altogether,91 compounds have been assigned to their originating plants and 84 substances were identified either by comparison with authentic references or with data from the literature.The HPTLC method is based on the application of two different mobile phases and is able to detect both lipophilic and hydrophilic constituents of m GQD.Conclusions:The modified GQD was extensively characterized by UHPLC combined with DAD and Q-Exactive Orbitrap high-resolution MS detection,leading to the assignment and identification of compounds present in the decoction.In addition,a new method for the fingerprint analysis of the m GQD using HPTLC was established,which allows fast and simple identification of the herbal ingredients in the mixture.展开更多
基金financially supported by the Austrian Federal Ministry of Education,Science and Research(402.000/00012-WF/V/6/2016),Vienna,Austriaby the China Academy of Chinese Medical Sciences as a“Belt and Road”special international cooperation project(GH2017-03-06),Beijing,Chinaby the Key Project of the National Natural Science Foundation(No.81430097),Beijing,China。
文摘Objective:Gegen Qinlian decoction(GQD)is a classical traditional Chinese medicine formulation which has been used for almost 2000 years.At Guang'anmen Hospital,Beijing,a modified GQD version(m GQD)with seven instead of four herbal ingredients has been applied to treat Type 2 diabetes.Quality control is a crucial prerequisite for the therapeutic application of herbal medicines.For the identification of products derived from classical GQD,the Chinese Pharmacopeia requires the analysis of only three marker compounds.Because m GQD is a more complex mixture containing seven herbs and hundreds of constituents,the pharmacopoeia method for GQD is inadequate.Materials and Methods:A more comprehensive characterization of the formula's constituents has been developed using ultra-high-performance liquid chromatography-diode array detection(UHPLC-DAD)-Q-Exactive-mass spectrometry(MS)in electrospray ionization positive and negative mode.Moreover,a new method for the fingerprint analysis of m GQD via high-performance thin-layer chromatography(HPTLC)has been established.Results:Altogether,91 compounds have been assigned to their originating plants and 84 substances were identified either by comparison with authentic references or with data from the literature.The HPTLC method is based on the application of two different mobile phases and is able to detect both lipophilic and hydrophilic constituents of m GQD.Conclusions:The modified GQD was extensively characterized by UHPLC combined with DAD and Q-Exactive Orbitrap high-resolution MS detection,leading to the assignment and identification of compounds present in the decoction.In addition,a new method for the fingerprint analysis of the m GQD using HPTLC was established,which allows fast and simple identification of the herbal ingredients in the mixture.
文摘目的利用分子对接和网络药理学的方法预测桂枝芍药知母汤治疗类风湿关节炎的主要活性成分和作用靶点,结合中医方解配伍理论对其多成分-多靶点-多通路的作用进行分析。方法在TCMSP、TCM-Datebase@Taiwan、PubChen Compound数据库收集桂枝芍药知母汤中9味中药(桂枝、芍药、知母、甘草、麻黄、生姜、白术、防风、附子)的主要化学成分。通过DrugBank、TTD数据库查找与类风湿关节炎治疗相关的蛋白靶点,并上传到String在线数据库,构建蛋白互作网络关系。在PDB数据库下载合适的蛋白靶点晶体结构,采用Discovery studio 4.5.0软件进行化合物与靶点的分子对接,利用Cytoscape 3.6.1软件,构建药物-化合物-靶点可视化网络,阐释桂枝芍药知母汤抗类风湿关节炎的主要作用机制。结果分子对接结果显示桂枝芍药知母汤中存在316个潜在的抗关节炎活性成分,作用于26个靶点,其中MAPK1、ZADH2、P38、AKR1C2、DHODH、CA2、MMP3、MMP9、RANKL等蛋白是主要作用靶点。生物功能和通路分析提示,桂枝芍药知母汤的作用机制主要涉及骨吸收(28%)、组蛋白激酶活性(20%)、肽基酪氨酸磷酸化(20%)、前列腺素代谢过程(12%)等生物过程。作用通路主要是破骨细胞的分化(94.12%)。结论采用分子对接结合网络药理学的方法,从多靶点多途径的角度研究桂枝芍药知母汤治疗类风湿关节炎的药效物质基础及分子机制,为更好的临床用药提供参考和依据。
