基于网络药理学和实验验证探讨小儿止哮平喘颗粒治疗儿童哮喘机制

Exploration on Mechanism of Xiaoer Zhixiao Pingchuan Granules in the Treatment of Pediatric Asthma Based on Network Pharmacology and Experimental Validation

目的基于网络药理学及动物实验验证探究小儿止哮平喘颗粒治疗儿童哮喘的潜在作用机制。方法检索TCMSP、BATMAN-TCM、UniProt数据库并筛选小儿止哮平喘颗粒的药物活性成分及其相关靶点,利用GeneCards、DisGeNET、OMIM数据库筛选儿童哮喘疾病靶点,将二者交集靶点通过STRING数据库获取靶点蛋白相互作用(PPI)关系,并导入Cytoscape3.8.0软件构建PPI网络,通过Metascape平台进行GO和KEGG通路富集分析,得出潜在作用通路。制备卵清蛋白致敏幼龄小鼠哮喘模型,予不同浓度小儿止哮平喘颗粒进行干预,通过HE染色和PAS染色评估组织病理学改变,通过Western blot验证网络药理学预测结果。结果筛选得到小儿止哮平喘颗粒活性成分154种,涉及儿童哮喘作用靶点283个。KEGG通路富集分析显示,交集靶点富集度高的信号通路有PI3K-Akt信号通路、TNF信号通路、MAPK信号通路等。活性成分-靶点-信号通路网络拓扑分析获得13个关键靶点。动物实验表明,小儿止哮平喘颗粒可明显改善卵清蛋白诱导的气道炎症和杯状细胞增生,并显著下调PI3K-Akt信号通路关键蛋白表达(P<0.05)。结论小儿止哮平喘颗粒治疗儿童哮喘通过多途径、多靶点发挥临床效用,其中PI3K-Akt信号通路可能是关键分子机制。

Objective To explore the potential mechanism underlying the treatment of pediatric asthma using Xiaoer Zhixiao Pingchuan Granules through network pharmacology analysis and animal experimental validation.Methods Active components and their associated targets in Xiaoer Zhixiao Pingchuan Granules were identified through screening and retrieval of TCMSP,BATMAN-TCM,and UniProt databases.Disease-related targets for pediatric asthma were selected from GeneCards,DisGeNET,and OMIM databases.The target protein-protein interaction(PPI)relationship between the intersecting targets of the two was obtained through the STRING database,and import it into Cytoscape 3.8.0 software to construct a PPI network.GO and KEGG enrichment analyses were conducted using the Metascape platform to identify potential pathways.An asthmatic mouse model was induced by ovalbumin,and different concentrations of Xiaoer Zhixiao Pingchuan Granules were administered as interventions.Histopathological changes were evaluated using HE staining and PAS staining,and the network pharmacology findings were validated through Western blot analysis.Results A total of 154 active ingredients targeting 283 pediatric asthma-related genes were identified in Xiaoer Zhixiao Pingchuan Granules.KEGG enrichment analyses revealed significant enrichment of signaling pathways such as the PI3K-Akt signaling pathway,TNF signaling pathway,and MAPK signaling pathway among intersection targets.Thirteen key targets were identified through topological analysis of ingredients-targets-pathways network.Animal experiments demonstrated that Xiaoer Zhixiao Pingchuan Granules significantly alleviated ovalbumin-induced airway inflammation and goblet cell hyperplasia,while downregulating the expression of key proteins in the PI3K-Akt signaling pathway(P<0.05).Conclusion The therapeutic efficacy of Xiaoer Zhixiao Pingchuan Granules in pediatric asthma involves a multi-pathway and multi-target mechanism,with the PI3K-Akt signaling pathway emerging as a potential key molecular target.