基于实验筛选和网络药理学研究二咖啡酰奎宁酸抗过敏性鼻炎的作用机制

Mechanism of Dicaffeoylquinic Acid Against Allergic Rhinitis Based on Experimental Screening and Network Pharmacology

关键词:  二咖啡酰奎宁酸  黑沙蒿  脱颗粒  过敏性鼻炎  网络药理学

Key words:  dicaffeoylquinic acid  Artemisia ordosica  degranulation  allergic rhinitis  network pharmacology

摘要:

目的 筛选黑沙蒿中有抗肥大细胞脱颗粒的二咖啡酰奎宁酸类化合物（dicaffeoylquinic acid，DCQAs），并结合网络药理学方法对活性化合物抗过敏性鼻炎作用机制进行探索。方法 通过检测细胞生存率、β-氨基己糖苷酶释放率及中性红染色建立C48/80诱导P815细胞脱颗粒的最佳方案；检测β-氨基己糖苷酶释放率筛选DCQAs；通过STITCH、Swiss、TCMID、TCMSP及GeneCards数据库收集活性化合物靶点、过敏性鼻炎和肥大细胞脱颗粒的相关靶点，获得共同有效靶点；应用String数据库和Cytoscape 3.7.2软件构建化合物-潜在有效靶点的网络图；利用DAVID数据库对有效靶点进行GO （Gene Ontology）功能富集分析和KEGG （Kyoto Encyclopedia of Genes and Genomes）通路富集分析。结果 6种DCQAs中，仅有3，5-DCQA和4，5-DCQA可显著减少肥大细胞脱颗粒；通过筛选发现18个潜在有效靶点，其中整合素β-1（integrin β-1，ITGB1）、中性粒细胞弹性蛋白酶（neutrophil elastase，ELANE）、72 kDa IV型胶原酶（72 kDa type IV collagenase，MMP2）、原癌基因酪氨酸蛋白激酶（proto-oncogene tyrosine-protein kinase Src，SRC）以及caspase-3可能是中心节点；GO和KEGG分析显示活性DCQAs通过白细胞跨内皮转移信号通路、GnRH信号通路和肿瘤信号通路等参与MAPK正向调节、细胞外基质分解和整合素介导的信号通路等生物学过程。结论 3，5-DCQA和4，5-DCQA为黑沙蒿提取物抗过敏性鼻炎的潜在活性化合物，通过多靶点-多途径发挥作用，为黑沙蒿提取物治疗过敏性鼻炎提供科学依据。

Abstract:

OBJECTIVE To screen the effect of dicaffeoylquinic acid compounds(DCQAs) from Artemisia ordosica for resistance to degranulation of mast cells and to explore the mechanism of the active compounds on allergic rhinitis by network pharmacology. METHODS The optimal protocols of degranulation in P815 induced by C48/80 was established by detecting cell viability, β-hexosaminidase release rate and the neutral red staining. The active DCQAs was screened by measuring the release rate of β-hexosaminidase. The target of the active DCQAs and the therapeutic targets for allergic rhinitis and mast cell degranulation were searched and collected from the database of STITCH, Swiss, TCMID, TCMSP, and GeneCards. Then the common effective targets were obtained. The network map of active DCQAs and the potential effective targets was conducted by the database of String and Cytoscape 3.7.2 software. GO(Gene Ontology) and KEGG(Kyoto Encyclopedia of Genes and Genomes) enrichment analysis were performed by DAVID database. RESULTS 3,5-DCQA and 4,5-DCQA could significantly reduce the release rate of β-hexosaminidase among the 6 kinds of DCQAs. Eighteen potential effective targets were collected and integrin β-1(ITGB1), neutrophil elastase(ELANE), 72 kDa type IV collagenase(MMP2), proto-oncogene tyrosine-protein kinase Src(SRC), and caspase-3 could be the hub proteins. GO and KEGG analysis showed that active DCQAs participated in the biological processes of MAPK positive regulation, extracellular matrix decomposition and integrin-mediated signaling pathways through leukocyte transendothelial metastasis signaling pathway GnRH signaling pathway, and tumor signaling pathway. CONCLUSION 3,5-DCQA and 4,5-DCQA are identified as potential active compounds of Artemisia ordosica extracts against allergic rhinitis, which exerted anti-allergic rhinitis effects through multiple targets-multiple pathways. It provids a scientific basis for the treatment of allergic rhinitis with Artemisia ordosica extracts.