基于指纹图谱和网络药理学的茯苓4个药用部位饮片质量标志物预测分析

Predictive Analysis of Quality Markers of Four Medicinal Parts of Poria cocos Decoction Pieces Based on Fingerprint and Network Pharmacology

目的基于指纹图谱、网络药理学及分子对接技术初步分析茯苓[Poria cocos(Schw.)Wolf]4个药用部位饮片(茯苓皮、赤茯苓、白茯苓、茯神)中的潜在质量标志物(quality marker,Q-Marker)并测定其含量。方法采用HPLC建立茯苓饮片指纹图谱,确认共有峰并进行指认,对18批不同药用部位茯苓饮片进行相似度评价、聚类分析和正交偏最小二乘-判别分析(OPLS-DA),结合网络药理学分析茯苓三萜酸成分的对应靶点和通路,构建“活性成分-靶点-通路”网络图,并进行了分子对接分析,预测茯苓饮片的Q-Marker并测定其在茯苓4个药用部位饮片中的含量。结果茯苓4个药用部位饮片HPLC指纹图谱中标定了15个共有峰,通过与对照品比对指认了其中6个色谱峰,分别为茯苓新酸B(2号峰)、去氢土莫酸(3号峰)、茯苓新酸A(4号峰)、去氢茯苓酸(9号峰)、茯苓酸(10号峰)和松苓新酸(12号峰);同一药用部位茯苓饮片相似度均大于0.97,不同药用部位的茯苓指纹图谱有明显差异,15个共有峰所代表成分的量在茯苓中根据药用部位由外向里(茯苓皮、赤茯苓、白茯苓和茯神)均呈现减少趋势。OPLS-DA筛选出茯苓4个药用部位饮片的7个潜在标志性色谱峰,其中包含5个指认的色谱峰,网络药理学分析表明,指认出的色谱峰所代表的三萜酸成分具有广泛的靶点和网络通路,可能与茯苓抗菌、抗炎、抗肿瘤等药理作用密切相关。分子对接结果与网络药理学结果一致。此外,4个药用部位茯苓饮片含有的6个三萜酸类成分中单一化合物的含量及其总质量分数(0.041%~0.495%)均是茯苓皮>赤茯苓>白茯苓和茯神。结论所建立的指纹图谱方法准确、可靠,同时结合网络药理学分析和分子对接技术预测了茯苓4个药用部位饮片中6个潜在Q-Marker的活性,为茯苓饮片质量的全面控制和评价提供了参考,也为茯苓的深度开发应用提供了依据。

OBJECTIVE To analyze the potential quality markers(Q-markers)of four medicinal parts of Poria cocos(Schw.)Wolf decoction pieces including Poria Cutis,Rubra Poria,white Poria and Poriacum Radix Pini Basedon HPLC fingerprint,network pharmacology and molecular dockingtechnology and determine their contents.METHODS HPLC method was used to establish the fingerprints of four medicinal parts of P.cocos decoction pieces,confirm common peaks and identify the major peaks.Eighteen batches of P.cocos decoction pieces were analyzed by using similarity evaluation,cluster analysis(CA)and orthogonal partial least squares discriminant analysis(OPLS-DA)methods.The targets and pathways of triterpenes in P.cocos were screened,and the network diagram of“component-target-pathway”was constructed by network pharmacology,and further predicated by molecular docking method,finally the potential Q-markers of P.cocos decoction pieces were analyzed and determined.RESULTS A total of 15 common peaks were demarcated in the fingerprints of four medicinal parts of P.cocos.Six of them were identified as poricoic acid B(peak 2),dehydrotumulosic acid(peak 3),poricoic acid A(peak 4),dehydropachymic acid(peak 9),pachymic acid(peak 10)and dehydrotrametenolic acid(peak 12).The similarity of P.cocos decoction pieces in the same medicinal part were greater than 0.97.And the fingerprints of different medicinal parts of P.cocos decoction pieces were obviously different.The contents of the 15 common peaks in P.cocos showed a decreasing trend from the outside to the inside of P.cocos referring to Poria Cutis,rubra Poria,white Poria and Poria cum Radix Pini.Seven potential chromatographic peaks of four medicinal parts of P.cocos decoction pieces were revealed by OPLS-DA,and five of them were identified chromatographic peaks which have a wide range of targets and network pathways and may be closely related to the antibacterial,anti-inflammatory and anti-tumor effects of P.cocos.The results of molecular docking were consistent with those of network pharmacology.In addition,the content of both single compound and their total mass fraction(0.041%~0.495%)of 6 triterpenes in four medicinal parts of P.cocos decoction pieces showed the same trend,indicating that the content of both single and total 6 verified triterpenes in Poria Cutis was higher than in rubra Poria,which was higher than that in white Poria and Poria cum Radix Pini.CONCLUSION The established fingerprint method is accurate and reliable,and the activities of six potential Q-markers in four medicinal parts of P.cocos decoction pieces are predicted,analyzed and verified by network pharmacology and molecular docking technique,which provides a reference for the comprehensive control and evaluation of P.cocos decoction pieces quality,and also provides a theoretical basis for its further development and application.