基于网络药理学的山豆根调控白细胞跨内皮迁移通路改善急性咽炎的作用机制

Mechanism of Sophora tonkinensis Gagnep regulating leukocyte transendothelial migration pathway in improving acute pharyngitis based on network pharmacology

目的基于网络药理学、动物实验及荧光定量PCR技术阐明山豆根改善急性咽炎的分子机制。方法利用中药系统药理学分析平台(Traditional Chinese Medicine systems pharmacology database and analysis platform,TCMSP)数据库收集山豆根的活性成分和作用靶点。通过GeneCards、OMIM、DrugBank和Disgenet数据库获取急性咽炎相关的靶点。筛选获得两者的共同靶标后,利用STRING数据库进行构建蛋白互作网络,采用Metascape平台进行通路分析。同时应用Cytoscape软件构建“中药-疾病-成分-靶点”网络和“中药-疾病-成分-靶点-通路”网络。建立大鼠急性咽炎模型,研究山豆根水提取物对大鼠急性咽炎的影响。利用荧光定量PCR技术研究山豆根对大鼠急性咽炎模型咽部组织关键通路上的关键基因靶点的影响。结果该实验获取山豆根活性成分21个的相关作用靶点509个,获取急性咽炎相关的靶点2167个,山豆根与急性咽炎的共同靶点194个。KEGG通路分析筛选了344条相关信号通路,其中显示IL-17信号通路、NF-kappa B信号通路和白细胞跨内皮迁移通路可能在山豆根改善急性咽炎的过程中起关键作用。动物实验表明,山豆根水提取物低剂量组对急性咽炎治疗效果较好。荧光定量PCR实验结果表明,山豆根低剂量组明显下调了白细胞跨内皮迁移通路ITGB2、PIK3CA、PIK3CD和PTPN11基因的表达水平(P<0.05)。结论山豆根改善急性咽炎具有多成分、多靶点、多通路协同作用的特点。

Aim To elucidate the molecular mechanism of Sophora tonkinensis Gagnep in improving acute pharyngitis based on network pharmacology,animal experiments and quantitative real-time PCR.Methods The active components and targets of Sophora tonkinensis Gagnep were collected from the database of traditional Chinese medicinal systems databases and analysis platform(TCMSP).Targets related to acute pharyngitis were acquired through GeneCards,OMIM,DrugBank and Disgenet databases.After the common targets of the two were screened,the STRING database was used to construct the protein interaction network,and the Metascape platform was used for pathway analysis.At the same time,Cytoscape software was used to construct a network of“herbal-disease-component-target”and“herbal-disease-component-target-pathway”network.The acute pharyngitis models in rats were established to study the effect of water extract of Sophora tonkinensis Gagnep on acute pharyngitis in rats.Quantitative real-time PCR technology was used to study the effect of Sophora tonkinensis Gagnep on key gene targets in key pathways of pharyngeal tissues in rats with acute pharyngitis.Results In this experiment,509 related targets of 21 active components of Sophora tonkinensis Gagnep were obtained,2167 related targets of acute pharyngitis were obtained,and 194 common targets of Sophora tonkinensis Gagnep and acute pharyngitis were obtained.KEGG pathway analysis screened 344 related signaling pathways,indicating that IL-17 signaling pathway,NF-kappa B signaling pathway and leukocyte transendothelial migration pathway might play a key role in the improvement of acute pharyngitis by Sophorae tonkinensis Gagnep.Animal experiments showed that the low dose group of Sophora tonkinensis Gagnep water extract had better therapeutic effect on acute pharyngitis.The results of quantitative real-time PCR showed that the low-dose group of Sophora tonkinensis Gagnep significantly down-regulated the expression levels of ITGB2,PIK3CA,PIK3CD and PTPN11 genes in leukocyte transendothelial migration pathway(P<0.05).Conclusions The above results show that Sophora tonkinensis Gagnep has the characteristics of multi-component,multi-target and multi-pathway synergy in improving acute pharyngitis,which provides a theoretical basis for further study on the complex mechanism of Sophora tonkinensis Gagnep in improving acute pharyngitis.