基于指纹图谱、多指标定量及网络药理学的大黄质量标志物预测分析

Predictive Analysis of Rhei Radix et Rhizoma Quality Markers Based on Fingerprint,Multi-indicator Quantification and Network Pharmacology

目的建立大黄HPLC指纹图谱及多指标定量分析方法,结合网络药理学预测大黄潜在质量标志物。方法采用HPLC对20批掌叶大黄样品中的没食子酸、儿茶素、表儿茶素等13种指标性成分进行含量测定,采用中药色谱指纹图谱相似度评价系统进行相似度分析,以13种指标成分含量进行主成分分析、正交偏最小二乘判别分析,采用网络药理学预测大黄相关成分的作用靶点和通路,构建成分-靶点-通路网络图并对核心靶点进行器官定位分析以预测大黄的潜在质量标志物。结果指纹图谱相似度在0.882~0.998,共确定了15个共有峰,大黄素甲醚、儿茶素、大黄素-1-O-葡萄糖苷、大黄素甲醚-8-O-葡萄糖苷、芦荟大黄素-8-O-葡萄糖苷是差异化合物,可用于鉴别和区分大黄质量。经筛选得到15个化合物,8个核心靶点,包括AKT1、TP53、JUN、HSP90AA1、CASP3、EGF、TNF、IL-6,涉及对氧化应激的反应、对脂多糖的反应、内肽酶活性,涉及脂质和动脉硬化、MAPK信号传导途径、AGE-RAGE信号通路在糖尿病并发症中的作用、非酒精性脂肪性肝病等通路,并构建成分-靶点-通路图,预测没食子酸、儿茶素、芦荟大黄素、大黄酸、大黄素等为大黄潜在的质量标志物。结论所建立的大黄指纹图谱特征性强,可用于控制其质量,特征谱和网络药理学分析预测了大黄的质量标志物,这为进一步研究大黄的作用机制提供参考。

OBJECTIVE To establish HPLC fingerprinting and multi-indicator quantitative analysis methods for Rhei Radix et Rhizoma and to predict potential quality markers of it in combination with network pharmacology.METHODS The content of 13 index components,including gallic acid,catechin and epicatechin,was determined in 20 batches of Rheum palmatum L.samples by HPLC,and the similarity analysis was performed by using the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System.The network pharmacology was used to predict the targets and pathways of Rhei Radix et Rhizoma-related components,construct a network diagram of components-targets-pathways and perform organ localization analysis on the core targets to predict the potential quality markers of Rhei Radix et Rhizoma.RESULTS The similarity of fingerprint profiles ranged from 0.882 to 0.998,and a total of 15 common peaks were identified.Emodin-3-methyl ether,catechin,emodin-1-O-glucoside,physcion-8-O-glucoside and aloe-emodin-8-O-glucoside were the differential compounds,which could be used to identify and differentiate Rhei Radix et Rhizome quality.The screening yielded 15 compounds with 8core targets including AKT1,TP53,JUN,HSP90AA1,CASP3,EGF,TNF,IL-6,involved in response to oxidative stress,response to lipopolysaccharide,endopeptidase activity,involved in lipid and atherosclerosis,MAPK signaling pathway,AGE-RAGE signaling pathway in diabetic complications pathways.Non-alcoholic fatty liver disease,and the construction of a component-target-pathway map.Prediction of gallic acid,catechin,aloe-emodin,rhein acid and emodin as potential quality markers of Rhei Radix et Rhizoma.CONCLUSION The established fingerprint profile of Rhei Radix et Rhizoma is highly characterized and can be used to control its quality.The characterization profile and network pharmacological analysis predict the quality markers of Rhei Radix et Rhizoma,which provide a reference for further research on the mechanism of action of Rhei Radix et Rhizoma.