基于数据挖掘和网络药理学方法探讨中医药治疗系统性红斑狼疮的用药规律和作用机制

Exploration of the medication rules and mechanism of traditional Chinese medicine in the treatment of systemic lupus erythematosus based on data mining and network pharmacology

目的:基于数据挖掘分析中医药治疗系统性红斑狼疮(systemic lupus erythematosus,SLE)的用药规律并筛选核心药对,通过网络药理学方法和分子对接技术探讨核心药对治疗SLE的作用机制。方法:收集2012年6月—2022年12月在安徽中医药大学第一附属医院风湿病科住院治疗的SLE患者的资料进行分析,共获得472名患者的完整处方,运用Excel 2016对用药规律进行统计分析,通过IBM SPSS Modeler 18.0软件对中药进行关联规则分析;通过TCMSP数据库检索药物活性成分和靶点,以“systemic lupus erythematosus”为关键词,检索DrugBank、GeneCards、TTD、OMIM数据库获取疾病相关靶点,构建药物-疾病交集靶点韦恩图;采用Cytoscape 3.7.1软件构建蛋白相互作用网络图和“药物-活性成分-靶点”网络图,通过Hubba插件获得药物治疗SLE的核心靶点;通过Metascape数据库进行基因本体(Gene Ontology,GO)功能富集分析及京都基因及基因组百科全书(Kyoto Encyclopedia of Genes and Genome,KEGG)通路富集分析;利用AutoDock 4.2.6软件对核心靶点与主要成分进行分子对接,通过PyMOL 2.5.2进行可视化分析。结果:SLE患者益气健脾化湿(茯苓、山药、黄芪等)及滋阴清热(知母、青蒿、地骨皮等)中药使用频次较高,甘味药、寒性药以及归肝、脾、肾经的中药使用频次较高。知母-黄柏组合具有较高的支持度(41.31%)和置信度(71.79%),可能在SLE的治疗中发挥重要作用。网络药理学结果表明,知母-黄柏共有34个活性成分,97个潜在作用靶点,该药对治疗SLE的5个核心靶点包括肿瘤坏死因子(tumornecrosis factor,TNF)、蛋白激酶B(protein kinase B,Akt1)、白细胞介素-6(interleukin 6,IL-6)、白细胞介素-1β(interleukin1β,IL-1β)、肿瘤蛋白p53(tumor protein p53,TP53)。GO功能和KEGG通路富集分析结果显示,知母-黄柏组合治疗SLE与平滑肌适应、发热、丝氨酸蛋白酶抑制剂复合物等生物过程以及IL-17、TNF、MAPK等信号通路密切相关。分子对接显示核心靶点和活性成分具有良好的对接活性。结论:中医药治疗SLE以益气健脾化湿、滋阴清热药物为主,知母-黄柏是治疗SLE的关键药对,可能通过多成分、多靶点、多通路发挥对SLE的治疗作用。

Objective:To analyze the medication rules of traditional Chinese medicine in the treatment of systemic lupus erythematosus(SLE)and screen core couplet medicines based on data mining,and to explore the mechanism of core couplet medicines in the treatment of SLE by network pharmacology and molecular docking technology.Methods:The data of SLE patients hospitalized in the Department of Rheumatology of The First Affiliated Hospital of Anhui University of Chinese Medicine from June 2012 to December 2022 were collected and analyzed.A total of 472 patients′complete prescriptions were obtained.Excel 2016 was used for statistical analysis of medication rules,and IBM SPSS Modeler 18.0 software was used to analyze the association rules of Chinese herbs.Drug active ingredients and targets were searched in TCMSP database,and disease-related targets were searched in DrugBank,GeneCards,TTD and OMIM databases with“systemic lupus erythematosus”as the key word to construct drug-disease intersection target Venn diagram.Cytoscape 3.7.1 software was used to construct the protein-protein interaction network and the“drug-active ingredient-target”network.The core targets for drug treatment of SLE were obtained through the Hubba plugin.Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed by Metascape database.AutoDock 4.2.6 software was used to perform molecular docking between the core targets and the main components,and visualized analysis was performed by PyMOL 2.5.2.Results:The frequency of use of Chinese herbs for supplementing Qi,invigorating spleen,removing dampness(Fuling,Shanyao,Huangqi,etc.)and nourishing Yin and clearing heat(Zhimu,Qinghao,Digupi,etc.)for SLE patients was higher,and the frequency of use of Chinese herbs with sweet flavor,cold nature and channel tropism for liver,spleen and kidney channels was higher.The combination of Zhimu and Huangbai had high support(41.31%)and confidence(71.79%),and might play an important role in the treatment of SLE.The results of network pharmacology showed that there were 34 active ingredients and 97 potential targets in combination of Zhimu and Huangbai.There were 5 core targets of in the couplet medicines of Zhimu and Huangbai for SLE,including tumornecrosis factor(TNF),protein kinase B(protein kinase B,Akt1),interleukin-6(IL-6),interleukin-1β(IL-1β),and tumor protein p53(TP53).The results of GO function and KEGG pathway enrichment analysis showed that the treatment of SLE with the combination of Zhimu and Huangbai was closely related to the biological processes such as smooth muscle adaptation,fever,serine protease inhibitor complex,and IL-17,TNF,MAPK signaling pathways.Molecular docking showed good docking activity of the core targets and active components.Conclusion:In the treatment of SLE with traditional Chinese medicine,Chinese herbs are mainly used to invigorate Qi,invigorate spleen,remove dampness,nourish Yin and clear heat.As key couplet medicines for the treatment of SLE,Zhimu and Huangbai may play a role in the treatment of SLE through multi-component,multi-target and multi-pathway.