麻黄-苦杏仁药对治疗支气管哮喘机制研究

Study on the Mechanism of Mahuang(Ephedrae Herba)-Xingren(Armeniacae Semen Amarum)in the Treatment of Bronchial Asthma

目的运用网络药理学和分子对接的方法研究麻黄-苦杏仁药对治疗支气管哮喘的分子机制。方法利用中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)查找麻黄、苦杏仁的有效成分及相关靶点,并运用Uniprot数据库对靶点进行人类基因名称标注并规范化处理。通过OMIM及GeneCards查找支气管哮喘疾病靶点。将中药的靶点与疾病靶点进行比对,找出药物与疾病的共同蛋白靶点。运用Cytoscape3.3.0构建"成分-靶点"网络,找出麻黄-苦杏仁的主要活性成分。运用String数据库对共同靶点进行蛋白相互作用网络(protein-protein interaction,PPI)分析,找出关键靶点蛋白。运用Bioconductor对共同蛋白靶点进行基因本体论(gene ontology,GO)功能富集和京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析,预测麻黄-苦杏仁药对治疗疾病的作用机制。通过iGemdock及AutoDockTools软件对筛选出的主要活性成分与关键蛋白靶点的最优结合模式进行分子对接。结果筛选出麻黄-苦杏仁药对治疗支气管哮喘疾病的36个有效成分和80个有效靶点。其中主要活性成分8个,包括quercetin、kaempferol、luteolin、Stigmasterol、beta-sitosterol、Glabridin、(+)-catechin、naringenin。治疗疾病的关键靶点蛋白7个,分别是TP53、IL6、EGFR、VEGFA、CASP3、ESR1、MAPK8。GO功能富集共得到1591个条目,其中生物过程(BP)占比最高,达89%。KEGG通路富集分析得到127条信号通路,涉及的主要通路为TNF signaling pathway、PI3K-Akt signaling pathway、p53 signaling pathway、C-type lectin receptor signaling pathway。分子对接显示麻黄-苦杏仁药对的8个主要有效成分与7个关键靶点的最优结合模式都能完美对接。结论本研究显示了麻黄-苦杏仁药对可能通过调节TNF、PI3K-Akt、p53等信号通路来达到抗炎,降低气道高反应性以及气道上皮细胞损伤修复的功能,从而治疗支气管哮喘,也为后期实验验证提供了理论依据。

Objective To study the molecular mechanism of Mahuang(Ephedrae Herba)-Xingren(Armeniacae Semen Amarum)on bronchial asthma by using network pharmacology and molecular docking. Methods TCMSP database was used to find the active components and related targets of Mahuang(Ephedrae Herba)-Xingren(Armeniacae Semen Amarum),and Uniprot database was used to label and normalize the targets. The targets of bronchial asthma were identified by OMIM and GeneCards databases. The target of traditional Chinese medicine was compared with that of disease to find out the common protein target of drug and disease. Cytoscape3.3.0 was used to construct the"component-target"network to find out the main active components of Mahuang(Ephedrae Herba)-Xingren(Armeniacae Semen Amarum). PPI analysis was conducted on common targets using String database to find out key target proteins. Bioconductor was used to conduct GO functional enrichment and KEGG pathway enrichment analysis on common protein targets to predict the mechanism of Mahuang(Ephedrae Herba)-Xingren(Armeniacae Semen Amarum)in the treatment of diseases. The software iGemdock and AutoDockTools were used to conduct molecular docking for the optimal binding mode between the main active components screened and the key protein targets. Results 36 effective components and 80 effective targets of Mahuang(Ephedrae Herba)-Xingren(Armeniacae Semen Amarum)for treating bronchial asthma were screened out. There were 8 main active ingredients,including quercetin,kaempferol,luteolin,stigmasterol,beta-sitosterol,glabridin,(+)-catechin,naringenin. There were 7 key target proteins in the treatment of diseases,namely TP53,IL6,EGFR,VEGFA,CASP3,ESR1 and MAPK8. A total of 1591 items were obtained through GO enrichment,among which the biological process(BP)accounted for the highest proportion,up to 89%. Enrichment analysis of KEGG pathway yielded 127 signaling pathways,including TNF signaling pathway,PI3 K-Akt signaling pathway,p53 signaling pathway,and C-type lectin receptor signaling pathway. The molecular docking showed that the eight main active components of Mahuang(Ephedrae Herba)-Xingren(Armeniacae Semen Amarum)drug pair were perfectly connected with the optimal binding mode of the seven key targets. Conclusion This study shows that Mahuang(Ephedrae Herba)-Xingren(Armeniacae Semen Amarum) may achieve anti-inflammatory effects,reduce airway hyperresponsiveness and repair of airway epithelial cell damage by regulating signal pathways such as TNF,PI3 K-Akt and p53,thus treating bronchial asthma,and providing theoretical basis for later experimental verification.