基于网络药理学及动物实验探讨抗敏止嗽颗粒治疗支气管哮喘的作用机制

Study on the mechanism of Kangmin Zhisou Granules in the treatment of bronchial asthma based on network pharmacology and animal experiments

目的利用网络药理学方法探讨抗敏止嗽颗粒治疗支气管哮喘的潜在作用机制,并结合动物实验加以验证。方法借助BATMAN-TCM数据库筛选抗敏止嗽颗粒的活性成分及对应靶点信息,通过GeneCards、OMIM数据库获得支气管哮喘的相关疾病靶点,将药物靶点与支气管哮喘靶点取交集后导入STRING数据库,建立PPI网络,采用Cytoscape 3.9.1软件构建"药物-活性成分-交集靶点"网络,筛选核心靶点,利用DAVID数据库对核心靶点进行GO功能和KEGG通路富集分析。制备卵蛋白诱导的哮喘小鼠模型,经抗敏止嗽颗粒干预后,观察小鼠肺组织病理学变化,采用ELISA法检测血清TNF-α、IL-6、IL-1β水平。结果得到抗敏止嗽颗粒活性成分240个,潜在作用靶点1364个,筛选得到TNF、IL-6、AKT1、ALB、IL-1β等11个核心靶点。GO功能富集分析结果显示,抗敏止嗽颗粒治疗支气管哮喘主要涉及蛋白质磷酸化的正调节、炎症反应的调节、脂多糖反应等生物进程及TNF、MAPK、IL-17等信号通路。动物实验显示,抗敏止嗽颗粒可降低血清TNF-α、IL-6、IL-1β水平(P<0.05),减少小鼠肺组织炎性细胞浸润,从而缓解哮喘症状。结论抗敏止嗽颗粒可能通过作用于TNF-α、IL-6、IL-1β等靶点发挥抗炎作用,进而缓解哮喘症状。

Objective To deeply explore the potential mechanism of Kangmin Zhisou Granules in the treatment of bronchial asthma through network pharmacology method;To verify it with animal experiments.Methods The active components and corresponding target information of Kangmin Zhisou Granules were screened with the help of BATMAN-TCM database,and the related disease targets of bronchial asthma were obtained through GeneCards and OMIM databases.The drug targets and bronchial asthma targets were intersected and imported String database was used to establish PPI network.Cytoscape 3.9.1 software was used to draw the network diagram of"Chinese materia medica-active components-intersection targets"and the core targets were screened.GO and KEGG enrichment analysis was performed on the core targets using DAVID database.A mouse model of asthma induced by ovalbumin was prepared.After the intervention of Kangmin Zhisou Granules,the pathological changes of mouse lung tissue were observed,and the contents of serum TNF-α,IL-6,IL-1βwere detected by ELISA.Results Totally 240 active components and 1364 potential targets were obtained from Kangmin Zhisou Granules.Tumor necrosis factor(TNF),interleukin-6(IL-6),protein kinase B(AKT1),albumin(ALB),interleukin 1-beta(IL-1β)and other 11 core targets were obtained after screening.The results of GO enrichment analysis showed that the treatment of bronchial asthma by Kangmin Zhisou Granules mainly involved the positive regulation of protein phosphorylation,the regulation of inflammatory response,lipopolysaccharide response and other biological processes,as well as TNF,activated protein kinase(MAPK),interleukin-17(IL-17)and other signaling pathways.Animal experiments confirmed that Kangmin Zhisou Granules could reduce the expression levels of TNF-α,IL-6 and IL-1βin serum(P<0.05),and reduce the infiltration of inflammatory cells in the lung tissue of mice,thereby relieving asthma symptoms.Conclusion Kangmin Zhisou Granules may exert anti-inflammatory effects by acting on TNF-α,IL-6,IL-1βand other targets to alleviate asthma symptoms.