The Network Pharmacological Study and Experimental Verification of Huanglong Antitussive Granule in the Treatment of Asthma
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摘要:
目的 结合网络药理学方法和实验验证探讨黄龙止咳颗粒治疗哮喘的作用靶点及潜在机制。 方法 借助TCMSP、TCMID、Swiss Target Prediction等数据库检索黄龙止咳颗粒的化学成分和作用靶点。通过GeneCards、OMIM、Disgenet、Drugbank数据库筛选出哮喘的疾病靶点。使用Cytoscape软件构建“药物-成分-靶点-疾病”的网络图和潜在靶点的相互作用关系, 通过富集分析预测作用机制。构建哮喘小鼠模型, 通过病理染色, ELISA检测, qPCR及Western blot来验证网络药理学富集分析结果。 结果 共获得黄龙止咳颗粒中175种活性成分、1 026个药物靶点, 哮喘900个疾病靶点, 得到药物-疾病共同靶点153个, GO富集分析共得到条目1 967个, KEGG通路富集筛选出包括TNF信号通路在内的共计84条信号通路。动物实验表明, 黄龙止咳颗粒能有效改善哮喘小鼠的气道炎症, 并通过影响TNF信号通路从而抑制MAPK和NF-κB信号通路。 结论 该研究初步揭示了黄龙止咳颗粒治疗哮喘的作用机制, 为临床用药和后续研究提供参考依据。 Abstract:OBJECTIVE Combining network pharmacology methods and experimental verification to explore the target and potential mechanism of Huanglong Antitussive Granule in treating asthma. METHODS With the help of TCMSP, TCMID, Swiss Target Prediction and other databases, the chemical components and targets of Huanglong Antitussive Granule are searched. Through GeneCards, OMIM, Disgenet, Drugbank database to screen out the disease targets of asthma. Use Cytoscape software to construct a network diagram of "drug-component-target-disease" and the interaction relationship between potential targets, and predict the mechanism of action through enrichment analysis. Mice model of asthma was constructed, and the results of network pharmacology enrichment analysis were verified by pathological staining, Elisa detection, q-PCR and Western Blot. RESULTS A total of 175 active ingredients, 1 026 drug targets, 900 disease targets for asthma and 153 common drug-disease targets were obtained in Huanglong Antitussive Granule. A total of 1 967 entries were obtained by GO enrichment analysis, and KEGG pathway enrichment screened out a total of 84 signaling pathways including TNF signaling pathways. Animal experiments show that Huanglong Antitussive Granule can effectively improve airway inflammation in asthmatic mice, and inhibit the MAPK and NF-κB signaling pathways by affecting the TNF signaling pathway. CONCLUSION This study initially revealed the mechanism of action of Huanglong Antitussive Granule in treating asthma, providing a reference for clinical medication and follow-up research. -
Key words:
- Huanglong Antitussive Granule /
- asthma /
- network pharmacology /
- molecular mechanism
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表 1 引物序列
Table 1. Primer sequence
基因名 上游引物 下游引物 IL-6 CTCCCAACAGACCTGTCTATAC CCATTGCACAACTCTTTTCTCA TNF ATGTCTCAGCCTCTTCTCATTC CGATCACCCCGAAGTTCAGTAG MAPK3 CAGCTCAACCACATTCTAGGTA TCAAGAGCTTTGGAGTCAGATT PTGS2 ATTCCAAACCAGCAGACTCATA CTTGAGTTTGAAGTGGTAACCG IL-1β GCCAGTGAAATGATGGCTTATT AGGAGCACTTCATCTGTTTAGG TLR4 GCCATCATTATGAGTGCCAATT AGGGATAAGAACGCTGAGAATT MMP9 CAAAGACCTGAAAACCTCCAAC GACTGCTTCTCTCCCATCATC ICAM1 CTGAAAGATGAGCTCGAGAGTG AAACGAATACACGGTGATGGTA MPO CAGGAACAACATCACCATTCG CTTGGAAGCGTGTATTGATAGC GAPDH AGGTCGGTGTGAACGGATTTG GGGGTCGTTGATGGCAACA 表 2 中药-成分-靶点基本信息统计表
Table 2. Chinese medicines-components-targets statistics
药物名称 成分数量 预测靶点数量 地龙 6 186 黄芪 21 450 桔梗 8 278 麻黄 25 365 山楂 74 694 射干 17 445 葶苈子 12 262 淫羊藿 23 486 鱼腥草 7 278 -
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