摘要
目标采用网络药理学和分子对接技术预测分析金嗓开音胶囊治疗咽炎的活性成分及作用机制,并进行初步的体外验证。方法通过TCMSP、Swiss TargetPrediction、Uniprot等数据库和文献挖掘获取金嗓开音胶囊的活性成分与作用靶点,利用GeneCards等数据库检索咽炎疾病作用靶点,韦恩图分析获得活性成分与疾病靶点交集,应用STRING数据库和Cytoscape 3.9.1软件构建蛋白互作网络图,通过拓扑方法分析出关键靶点,对其进行GO和KEGG通路富集分析并构建可视化的“药物-成分-靶点-通路”网络图,筛选关键成分、靶点进行分子对接研究。最后,建立细胞炎性模型,采用MTT、ELISA、Western blot法对核心靶点进行验证。结果检索到金嗓开音胶囊活性成分共251个,靶点1101个,咽炎相关疾病靶点1459个,筛选出PTGS2、TNF、IL-6等核心靶点25个,槲皮素、木犀草素、β-谷甾醇等关键成分10个,涉及MAPK信号通路、TNF信号通路及IL-17信号通路等。分子对接结果表明,金嗓开音胶囊中的槲皮素、山柰酚、β-谷甾醇等主要活性成分与PTGS2、TNF、IL-6等关键靶点具有较强结合力,MTT、ELISA实验结果表明,金嗓开音胶囊对脂多糖诱导的巨噬细胞炎性模型具有一定的保护作用,并可抑制炎性因子NO、TNF-α及IL-6的表达,Western blot实验结果显示金嗓开音胶囊可抑制PTGS2、TNF-α及IL-6蛋白的表达。结论金嗓开音胶囊可能通过多成分、多靶点、多通路影响炎症和免疫调节,从而发挥治疗咽炎的作用。
Obiective To determine the mechanism of Jinsang Kaiyin capsules for pharyngitis by network pharmacology and molecular docking,and to validate in vitro experiments.Methods The active components and corresponding targets were obtained from TCMSP,Swiss TargetPrediction,Uniprot and other databases and literature mining.The targets of pharyngitis were searched with GeneCards and other databases,and the intersections of active components and disease targets were analysed by Venn diagram.The protein-protein interaction network between the active components and the disease targets was constructed with STRING database and Cytoscape 3.9.1 software.The core targets were identified by topological analysis.GO and KEGG pathway enrichment analysis were performed on the core targets,and the visualized“drug-ingredient-target-pathway”network map was constructed.The core components and targets were screened for molecular docking.Finally,cellular inflammation models were established and the core targets were validated by MTT,ELISA and Western blot.Results Totally 251 active components,1101 targets and 1459 pharyngitis-related disease targets were retrieved.Totally 25 core targets(such as PTGS2,TNF and IL-6)and 10 core components(such as quercetin,luteolin andβ-sitosterol)were screened,involving MAPK signaling pathway,TNF signaling pathway and IL-17 signaling pathway,etc.The molecular docking showed that the main active components in Jinsang Kaiyin capsules,such as quercetin,kaempferol andβ-sitosterol,had stable binding power to core targets such as PTGS2,TNF and IL-6.The MTT and ELISA results showed that lipopolysaccharide-induced macrophage inflammation model was improved by Jinsang Kaiyin capsules,and the expression levels of cytokines NO,TNF-αand IL-6 were significantly decreased by Jinsang Kaiyin capsules.Western blot results showed that Jinsang Kaiyin capsules inhibited the expression of PTGS2,TNF-αand IL-6 protein.Conclusion Jinsang Kaiyin capsules are effective for pharyngitis via multiple components,targets and pathways to influence the inflammation and immunity regulate.
作者
任美
周瑞
唐志书
苏洁
刘妍如
胡锦航
宋忠兴
石丽
王文宏
REN Mei;ZHOU Rui;TANG Zhi-shu;SU Jie;LIU Yan-ru;HU Jin-hang;SONG Zhong-xing;SHI Li;WANG Wen-hong(Shaanxi University of Chinese Medicine Co-construction Collaborative Innovation Center of Chinese Medicine Resources Industrialization by Shaanxi&Education Ministry/State Key Laboratory of Research&Development of Characteristic Qin Medicine Resources(Cultivation)/Shaanxi Innovative Drug Research Center,Shaanxi University of Chinese Medicine,Xianyang Shaanxi 712083;Xi’an Beilin Pharmaceutical Co.,Ltd,Xi’an 712000)
出处
《中南药学》
CAS
2022年第7期1550-1558,共9页
Central South Pharmacy
基金
国家自然科学基金青年项目(No.81803946)
陕西省教育厅2020年度重点科学研究计划(协同创新中心项目)(No.20JY010)
陕西省2020年重点研发计划社会发展领域(No.2020SF-034)
国家中药材产业技术体系咸阳综合试验站补贴项目(No.2019JCW-06)。
关键词
金嗓开音胶囊
咽炎
网络药理学
分子对接
细胞实验
Jinsang Kaiyin capsule
pharyngitis
network pharmacology
molecular docking
cell experiment
