基于指纹图谱、化学计量学、网络药理学的半夏汤洗前后质量评价

Quality evaluation of Pinelliae Rhizoma before and after hot water washing based on HPLC fingerprint,chemometrics and network pharmacology

目的建立半夏汤洗前后HPLC指纹图谱及化学模式识别方法,并利用网络药理学方法分析半夏药理作用的潜在靶点和作用机制。方法采用Waters C_(18) Xselect(250 mm×4.6 mm,5μm)色谱柱,以乙腈-水为流动相,梯度洗脱,体积流量1.0 mL/min,柱温30℃,检测波长260 nm,用相似度评价、主成分分析(PCA)、正交偏最小二乘法-判别分析(OPLS-DA)对汤洗前后半夏质量进行分析。结合网络药理学分析,利用Swiss Target Prediction和David等数据库分析半夏成分的对应靶点和通路,并在Cytoscape 3.6.0软件中绘制出"成分-靶点-通路"网络图。结果半夏药材指纹图谱中标定了20个共有峰,通过与对照品比对,指认了其中7个色谱峰,分别为次黄嘌呤(7号峰)、黄嘌呤(8号峰)、尿苷(9号峰)、腺嘌呤(10号峰)、胸苷(13号峰)、腺苷(14号峰)和琥珀酸(20号峰),13批半夏药材相似度均大于0.95,PCA显示S6批样品(甘肃陇南)综合质量较好。PCA和OPLS-DA结果表明半夏汤洗前后在化学成分组成和含量上有差异,且可明显区分,并揭示了贡献最大的4个潜在标志性色谱峰,分别为11、10(腺苷)、18、20(琥珀酸)号峰,其为导致差异的主要标志物。指认出的色谱峰具有广泛的靶点和网络通路,与半夏的主要药理作用密切相关。结论所建立的指纹图谱方法准确、可靠,可以有效地区分半夏和汤洗半夏,并为半夏功效关联物质的研究及作用机制的阐释奠定基础。

Objective To establish an HPLC fingerprint and chemical pattern recognition of Banxia(Pinelliae Rhizoma,PR)before and after hot water washing(wPR),and analyze the potential target and mechanism of PR by network pharmacology.Methods Waters C_(18) Xselect(250 mm×4.6 mm,5μm)column was used with acetonitrile-water as mobile phase for gradient elution with flow rate of 1.0 mL/min,column temperature of 30℃,detection wavelength of 260 nm.Similarity evaluation,principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were used for quality analysis of PR and wPR.Combined with network pharmacology analysis,Swiss target prediction and David database were used to analyze the corresponding target and pathway of components,and the"component-target-pathway"diagram was drawn in Cytoscape 3.6.0 software.Results A total of 20 common peaks were demarcated in the fingerprint of PR.Seven of them were identified as hypoxanthine(peak 7),xanthine(peak 8),uridine(peak 9),adenine(peak 10),thymidine(peak 13),adenosine(peak 14)and succinic acid(peak 20).The similarity of 13 batches of PR were greater than 0.95.PCA showed that the comprehensive quality of S6 batches of samples(Longnan,Gansu)was better.The results of PCA and OPLS-DA showed that there were differences in chemical composition and content between PR and wPR.Four potential chromatographic peaks,11,10(adenosine),18,20(succinic acid),which were the main markers leading to the difference,were revealed.The identified chromatographic peaks had a wide range of targets and network pathways,which were closely related to the main pharmacological effects of PR.Conclusion The fingerprints were accurate and reliable,which could effectively distinguish PR and w PR,and lay a foundation for the study of related substances of PR and the interpretation of its mechanism of action.