基于数据挖掘和网络药理学技术的《中医方剂大辞典》含连翘组方规律及其抗炎机制分析

Compatibility Rules of Prescriptions Containing Forsythiae Fructus in Dictionary of Traditional Chinese Medicine Prescriptions and Anti-inflammatory Mechanism:An Exploration Based on Data Mining and Network Pharmacology

目的:运用数据挖掘技术分析连翘组方配伍规律,并基于网络药理学方法分析连翘抗炎的药理机制,将对指导连翘中医临床合理应用、保健食品的开发及中药新药研发提供参考。方法:收集整理《中医方剂大辞典》中含连翘的方剂,构建临证方药数据库,并运用SPSS Statistics 26和IBM SPSS Modeler 18.0及Gephi 9.2软件对其配伍用药、治疗病症进行频次统计和关联规则、复杂网络等数据挖掘。利用中药系统药理学数据库与分析平台(TCMSP)数据库和BATMAN-TCM,SEA数据库收集连翘的活性成分及其抗炎作用靶点,通过GeneCards,人类孟德尔遗传数据库(OMIM),CTD,GenCLiP3数据库获取并筛选与抗炎相关的靶点。运用STRING平台分析蛋白质相互作用,构建蛋白质-蛋白质相互作用(PPI)网络。采用Metascape平台分析进行富集分析,并采用Cytoscape 3.8.2软件构建连翘活性成分-抗炎靶点-信号通路网络。结果:严格按照纳入与排除标准进行方剂筛选后提取方剂数据,收集含连翘方剂共2245首,涉及中药512味,用药总频次达27314次,其中与连翘相配伍的高频药物(>800次)为甘草(1483次),黄芩(964次),当归(842次),并选取连翘-黄芩和连翘-当归药对进行深度挖掘。含连翘组方主治疾病共29类,并筛选出其中具有代表性的"痈疽、疮疡""眼科病证"和"天行"三类高频主治病症进行数据挖掘,其中"痈疽、疮疡"病证得到以连翘-甘草-金银花-当归为核心药物组合的关联规则数据19条,聚类分析得到1组多味药物聚合组和4组药对及单味药金银花,复杂网络分析得到4组药物模块,因子分析得到6个公因子;"眼科病证"得到以连翘-甘草-黄芩-当归为核心药物组合的关联规则数据23条,聚类分析得到2组多味药物聚合组,4组药对,复杂网络分析得到4组药物模块,因子分析得到5个公因子;"天行"病证得到以连翘-甘草-薄荷-金银花为核心药物组合的关联规则数据28条,聚类分析得到3组多味药物聚合组、1组药对及单味药连翘、甘草,复杂网络分析得到4组药物模块,因子分析得到5个公因子。网络药理学结果显示,连翘抗炎的核心活性成分为槲皮素、木犀草素、山柰酚等,核心靶点有磷脂酰肌醇-3激酶调节亚基1(PIK3R1),蛋白激酶B1(Akt1),表皮生长因子受体(EGFR)等,生物学通路主要作用于proteoglycans in cancer,pathways in cancer,PI3K/Akt signaling pathway等信号通路,其功能主要为抑制转录因子、调节酶活性和炎症相关基因表达等。结论:该研究运用多种数据挖掘技术探讨连翘高频病证的组方配伍规律,客观、直观、科学地展现了连翘的配伍规律。针对不同病证,连翘皆常配伍清热药、补虚药、解表药、活血化瘀药进行治疗。在清热解毒的同时,强调扶助正气、疏通壅滞的重要性,而针对里热炽盛的程度不同,配伍以不同功效的清热药物。研究体现了连翘在具体病证中发挥的独特优势,揭示了连翘抗炎的多成分、多靶点、多通路的作用机制,突破了连翘在现代中医药临床及实验研究中的局限性,对指导连翘中医临床合理应用、保健食品的开发及中药新药研发具有重要意义,并有助于推动我国中医药大健康产业的发展。

Objective:To analyze the compatibility rules of prescriptions containing Forsythiae Fructus based on data mining and explore the anti-inflammatory mechanism of Forsythiae Fructus based on network pharmacology,so as to provide reference for the rational clinical application of Forsythiae Fructus and the development of health foods and new Chinese medicines. Method: The prescriptions containing Forsythiae Fructus in the Dictionary of Traditional Chinese Medicine Prescriptions were collected,based on which a clinical prescription database was constructed. The Chinese herbs combined with Forsythiae Fructus and the corresponding indications were subjected to frequency statistics,association rule analysis,and complex network analysis using SPSS Statistics 26,IBM SPSS Modeler 18.0,and Gephi 9.2. The active components and targets of Forsythiae Fructus for anti-inflammation were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),BATMAN-TCM,and SEA,and the targets related to anti-inflammation from GeneCards,Online Mendelian Inheritance in Man(OMIM),CTD,and GenCLiP3.Following the analysis of protein-protein interactions(PPI)with STRING,a PPI network was constructed. The enrichment analysis was performed using Metascape,and the active component-anti-inflammation targetsignaling pathway network of Forsythiae Fructus was constructed by Cytoscape 3.8.2. Result:According to the inclusion and exclusion criteria,2 245 prescriptions containing Forsythiae Fructus were harvested,involving 512 Chinese herbs,with a total usage frequency of 27 314. The Chinese herbs that were most frequently combined with Forsythiae Fructus( >800 times)were Glycyrrhizae Radix et Rhizoma(1 483 times),Scutellariae Radix(964 times),and Angelicae Sinensis Radix(842 times). Hence,the herbal pairs "Forsythiae Fructus-Scutellariae Radix" and "Forsythiae Fructus-Angelicae Sinensis Radix" were further explored. The prescriptions containing Forsythiae Fructus could be utilized for the treatment of 29 kinds of diseases,and three representative disease categories including "carbuncle,gangrene,sores and ulcers", "ophthalmic diseases and syndromes" and "epidemic diseases" are selected for data mining. There were 19 association rules obtained with "Forsythiae Fructus-Glycyrrhizae Radix et Rhizoma-Lonicerae Japonicae Flos-Angelicae Sinensis Radix" as the core herb combination for "carbuncle,gangrene,sores and ulcers". The clustering analysis revealed one multi-herb clustering group,four herbal pairs,and single herb Lonicerae Japonicae Flos,the complex network analysis four herbal modules,and the factor analysis six common factors. There were 23 association rules obtained with "Forsythiae Fructus-Glycyrrhizae Radix et Rhizoma-Scutellariae Radix-Angelicae Sinensis Radix" as the core herb combination for "ophthalmic diseases and syndromes". The clustering analysis revealed two multi-herb clustering groups and four herbal pairs,the complex network analysis four herbal modules,and the factor analysis five common factors. There were 28 association rules obtained with "Forsythiae Fructus-Glycyrrhizae Radix et Rhizoma-Menthae Haplocalycis Herba-Lonicerae Japonicae Flos" as the core herb combination for "epidemic diseases". The clustering analysis revealed three multi-herb clustering groups,one herbal pair,and two single herbs Forsythiae Fructus and Glycyrrhizae Radix et Rhizoma,the complex network analysis four herbal modules,and the factor analysis five common factors. As demonstrated by network pharmacology-based analysis,the core anti-inflammation components of Forsythiae Fructus were quercetin,luteolin,and kaempferol,and the core targets were phosphoinositide-3-kinase regulatory subunit 1(PIK3 R1),protein kinase B 1(Akt1),and epidermal growth factor receptor(EGFR). The biological pathways were mainly concentrated in proteoglycans in cancer,pathways in cancer,and PI3 K/Akt signaling pathway,with such functions as inhibition of transcription factors,regulation of enzyme activity,and inflammation-related gene expression involved. Conclusion: This study employed a variety of data mining techniques to objectively,intuitively,and scientifically uncover the compatibility rules of Forsythiae Fructus in the treatment of high-frequency diseases. It has been found that Forsythiae Fructus is often combined with heat-clearing herbs,tonifying herbs,exterior-releasing herbs,and blood-activating and stasis-resolving herbs for diverse diseases and syndromes. Under the premise of clearing heat and removing toxin,reinforcing healthy Qi and dredging stagnation are also emphasized. According to the degree of internal heat exuberance,the heat-clearing herbs with different merits are combined. This study has revealed the unique advantages of Forsythiae Fructus in the treatment of specific diseases and syndromes as well as its multi-component,multi-target,and multi-pathway mechanisms in anti-inflammation,breaking through the limitations in modern clinical and experimental research of Forsythiae Fructus. These findings are of great significance for guiding the rational clinical application of Forsythiae Fructus and the development of health foods and new Chinese medicines,thus better accelerating the development of Chinese medicine health industry.