指纹图谱结合网络药理学及分子对接的黑顺片质量标志物预测

Prediction of Quality Markers of Heishun Tablet Based on Fingerprint,Network Pharmacology and Molecular Docking

目的:结合指纹图谱、网络药理学和分子对接技术,初步分析黑顺片中的潜在质量标志物(Q-Marker)。方法:运用高效液相色谱法(HPLC)建立黑顺片饮片指纹图谱,确认共有峰并进行指认,同时进行聚类分析及主成分分析,运用网络药理学方法构建“活性成分-靶点-通路”网络,预测Q-Marker,采用分子对接验证。结果:建立了25批黑顺片饮片指纹图谱,确认了8个共有峰,通过对照品指认6个色谱峰,分别为苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、新乌头碱、次乌头碱、乌头碱,指纹图谱相似度为0.922~0.999。聚类分析将25批样品大致聚为2类,主成分分析显示S4、S10、S11批次的样品质量显著区别于其他批次;网络药理学分析表明,次乌头碱等6种成分均是黑顺片发挥药效的主要活性成分,主要发挥抗肿瘤、抗炎、保护心血管等作用,分子对接表明以上成分与关键靶点均有较强的结合活性,可能是黑顺片潜在Q-Marker。结论:本实验从多层面综合获取黑顺片的质量信息,对其质量标志物进行初步预测及验证,为进一步研究黑顺片作用机制,全面控制饮片质量提供参考。

Objective:To preliminarily analyze the potential quality markers(Q-Markers)of Heishun Tablet by combining fingerprint,network pharmacology and molecular docking.Methods:High performance liquid chromatography(HPLC)was used to establish the fingerprint of Heishun Tablet and identify the common peaks,and cluster analysis and principal component analysis were performed at the same time.Network pharmacology was employed to construct the“active component-target-pathway”network to predict Q-markers,and molecular docking was used for verification.Results:The fingerprints of 25 batches of Heishun Tablet were established.Eight common peaks were calibrated and 6 of them were identified,which were benzoylmesaconine,benzoylaconine,benzoylaconine,mesaconitine,hypaconitine and aconitine.The similarity ranged from 0.922 to 0.999.Cluster analysis showed that the 25 batches of samples were roughly clustered into two groups,and principal component analysis found that S4,S10 and S11 samples were significantly different from the other batches.Network pharmacology found that six components,including hypoaconitine,were the main active components of Heishun Tablet,and mainly played anti-tumor,anti-inflammatory,and cardiovascular protective functions.By molecular docking,the above components all had strong binding activity with the key targets,which might be the potential Q-markers of Heishun Tablet.Conclusion:In this paper,the quality information of Heishun Tablet was obtained from multiple levels,and the Q-markers were preliminarily predicted and verified.It is expected to facilitate the further research on the mechanism of Heishun Tablet and the overall quality control of decoction pieces.