辣椒风湿膏治疗关节痛作用机制的网络药理学研究

Mechanism of Lajiao Fengshi Plaster in the Treatment of Arthralgia Based on Network Pharmacology

目的探讨辣椒风湿膏治疗关节痛的作用机制。方法检索中药系统药理学数据库与分析平台(TCMSP),并结合PubMed数据库获得制剂组方药材的活性成分,通过SwissTargetPrediction数据库预测其靶点;通过GeneCards、OMIM、TTD数据库预测关节痛疾病靶点,利用Venn在线绘图软件筛选活性成分与疾病的共有靶点;将共有靶点上传至STRING数据库进行蛋白相互作用(PPI)分析;基于Metascape平台对共有靶点进行基因本体论(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析;借助Cytoscape 3.9.0软件构建疾病-活性成分-共有靶点-通路网络,利用该软件的Network Analyzer工具筛选辣椒风湿膏治疗关节痛的主要活性成分和核心靶点,并进行分子对接验证。结果共获得活性成分28个及其靶点440个,疾病靶点1296个,共有靶点105个。GO功能富集分析共获得生物功能条目1374条,其中生物过程1184条,细胞组分61条,分子功能129条;KEGG通路富集分共获得135条信号通路,主要涉及钙信号通路、环磷酸腺苷(cAMP)信号通路、瞬时受体电位(TRP)通道的炎性介质调节信号通路等。共获得主要活性成分6个(齐墩果酸、辣椒素、二氢辣椒素、辣椒素酯、左旋龙脑、薄荷醇)与核心靶点15个(mTOR、ESR1、TP53等)。分子对接结果显示,TRPV1、ESR1、PTGS2、mTOR和MMP-9靶点蛋白与除辣椒素酯外的上述其余5个主要活性成分结合活性良好(结合能<-1.2 kcal/mol)。结论辣椒风湿膏可能通过辣椒素、二氢辣椒素、齐墩果酸、左旋龙脑、薄荷醇等化学成分调控mTOR、ESR1、PTGS2、TP53、TRPV1等基因的表达,进而调控钙信号通路、cAMP信号通路等治疗关节痛。

Objective To investigate the mechanism of Lajiao Fengshi Plaster in the treatment of arthralgia.Methods The active ingredients of medicinal herbs in the preparation were obtained by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and PubMed database,the targets were obtained by the SwissTargetPrediction database.The targets of arthralgia were predicted by the GeneCards,OMIM,and TTD databases,the common targets of active ingredients and disease were screened by the Venn online plotting software,and the common targets were imported into the STRING database for protein-protein interaction(PPI)analysis.The Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis on common targets were performed based on the Metascape platform.The disease-active ingredient-common target-pathway network was constructed by the Cytoscape 3.9.0 software,the main active ingredients and core targets of Lajiao Fengshi Plaster in the treatment of arthralgia were screened by the Network Analyzer tool of this software,and the molecular docking was performed for verification.Results A total of 28 active ingredients,440 targets of active ingredients,1296 disease targets,105 common targets were obtained.The GO functional enrichment analysis showed 1374 biological functions,including 1184 biological processes,61 cellular components and 129 molecular functions.The KEGG pathway enrichment analysis showed 135 signaling pathways,mainly involving the calcium signaling pathway,cyclic adenosine monophosphate(cAMP)signaling pathway,and inflammatory mediator regulation of transient receptor potential(TRP)channels.Six main active ingredients[oleanolic acid,capsaicin,dihydrocapsaicin,capsiate,(-)-borneol,menthol]and 15 core targets(mTOR,ESR1,TP53,etc.)were obtained.The molecular docking showed that the TRPV1,ESR1,PTGS2,mTOR and MMP-9 target proteins had good binding activity with the above five main active ingredients except for capsiate,with the binding energy lower than-1.2 kcal/mol.Conclusion Lajiao Fengshi Plaster may regulate the expressions of mTOR,ESR1,PTGS2,TP53,TRPV1 and other genes through chemical components such as capsaicin,dihydrocapsaicin,oleanolic acid,(-)-borneol and menthol,and then regulate the calcium signaling pathway,cAMP signaling pathway to treat arthralgia.