基于UPLC-Q-TOF/MS代谢组学和分子对接技术探索胆黄连“清肝胆实火”的物质基础  

Exploration of Material Basis of"Clearing Excessive Fire of Liver-gallbladder"of Bile-processed Coptidis Rhizoma Based on UPLC-Q-TOF/MS Metabolomics and Molecular Docking Technology

作　　者：董兆威 田茂颖 杨宇[1] 王瑾[1] 黄勤挽[1] DONG Zhaowei;TIAN Maoying;YANG Yu;WANG Jin;HUANG Qinwan(State Key Laboratory of Southwestern Chinese Medicine Resources,School of Pharmacy,School of Ethnic Medicine,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

机构地区：[1]成都中医药大学西南特色中药资源国家重点实验室,药学院,民族医药学院,成都611137

出　　处：《中国实验方剂学杂志》2023年第23期140-149,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82173978)。

摘　　要：目的:采用超高效液相色谱-四极杆-飞行时间串联质谱法(UPLC-Q-TOF/MS)代谢组学及分子对接技术研究胆黄连清肝胆实火的物质基础。方法:采用UPLC-Q-TOF/MS代谢组学技术分析黄连、清炒黄连和胆黄连的整体化学成分,流动相0.1%甲酸水溶液(A)-乙腈(B)梯度洗脱(0~2 min,5%B;2~20 min,5%~65%B;20~40 min,65%~10%B;40~45 min,10%B;45~46 min,10%~95%B;46~49 min,95%B),电喷雾离子源(ESI),正、负离子模式扫描,扫描范围m/z 80~1200。在此基础上,结合偏最小二乘法-判别分析(PLS-DA)和方差分析筛选不同黄连炮制品之间的差异化合物。利用网络药理学和分子对接技术验证差异化合物与肝胆实火证的关联程度。结果:共鉴定了33个化学成分,包括2个酚酸类、5个结合型胆汁酸类、26个生物碱类。PLS-DA和方差分析确认差异化合物16个,其中11个生物碱、5个结合型胆汁酸。在京都基因与基因组百科全书(KEGG)数据库进行通路富集分析得到差异成分与“肝胆实火”相关的疾病通路8条,“成分-靶点-通路”网络分析得到关键靶点磷脂酰肌醇4,5-二磷酸3-激酶催化亚基α亚型(PIK3CA);分子对接结果显示,11种生物碱成分与PIK3CA均具有良好的结合能力。结论:猪胆汁在胆黄连炮制中具有独特性,可以促进小檗红碱、二氢白屈菜红碱等11种生物碱的生成和溶出。结合分子对接结果和文献结果可认为小檗红碱、二氢白屈菜红碱、牛磺猪去氧胆酸等16个差异化合物是胆黄连清肝胆实火的物质基础。Objective:To explore the material basis of bile-processed Coptidis Rhizoma clearing excessive fire of liver-gallbladder based on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF/MS)metabolomics and molecular docking.Method:UPLC-Q-TOF/MS metabolomics was used to analyze the chemical constituents of Coptidis Rhizoma,water-processed Coptidis Rhizoma and bile-processed Coptidis Rhizoma.Chromatographic separation was achieved with 0.1%formic acid aqueous solution(A)-acetonitrile(B)as the mobile phase in gradient elution(0-2 min,5%B;2-20 min,5%-65%B;20-40 min,65%-10%B;40-45 min,10%B;45-46 min,10%-95%B;46-49 min,95%B),and electrospray ionization(ESI)was applied and operated in positive and negative ion modes,the acquisition range was m/z 80-1200.Based on this,partial least squares-discriminant analysis(PLS-DA)and variance analysis were used to screen the differential compounds among the three products of Coptidis Rhizoma.Network pharmacology and molecular docking were used to verify the degree of association between differential compounds and excessive fire of liver-gallbladder syndrome.Result:A total of 33 chemical constituents were identified,including 2 phenolic acids,5 binding bile acids and 26 alkaloids.And 16 differential compounds were identified by multivariate statistical analysis,including 11 alkaloids and 5 binding bile acids.Pathway enrichment analysis in the Kyoto Encyclopedia of Genes and Genomes(KEGG)database yielded 8 pathways related to excessive fire of liver-gallbladder,and the key protein phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha isoform(PIK3CA)was obtained according to the"component-target-pathway"network analysis.Molecular docking results showed that 11 alkaloids had good binding ability with PIK3CA.Conclusion:Porcine bile is unique in the processing of bile-processed Coptidis Rhizoma,which can promote the production and dissolution of 11 alkaloids,including berberine and dihydrochelerythrine.Based on the results of molecula

关 键 词：胆黄连 肝胆实火 物质基础 代谢组学 分子对接技术 网络药理学 超高效液相色谱-四极杆-飞行时间串联质谱法(UPLC-Q-TOF/MS) 

分 类 号：R22[医药卫生—中医基础理论] R28[医药卫生—中医学] R943.1[理学—分析化学] O657[理学—化学]