基于网络药理学的三黄汤治疗湿疹作用机制研究

Study on the mechanism of Sanhuang decoction in the treatment of eczema based on network pharmacology

目的:基于网络药理学方法并结合体外THP-1巨噬细胞炎症模型预测三黄汤治疗湿疹的有效成分及可能作用机制。方法:采用中药系统药理数据库和分析平台(TCMSP)筛选三黄汤中的活性化学成分,通过GeneCard数据库查找化合物与湿疹共有靶点;采用Cytoscape软件构建“成分-靶点-疾病”网络,利用STRING数据库进行蛋白互作(PPI)网络分析,用DAVID数据库进行基因功能注释分析(GO)和信号通路富集分析(KEGG),预测三黄汤治疗湿疹的作用机制。使用PMA联合脂多糖(LPS)刺激THP-1细胞,同时三黄汤干预,荧光定量(qRT-PCR)、酶联免疫吸附(Elisa)分别检测细胞炎症因子表达,蛋白质印迹法(Western blot)检测相关蛋白表达。结果:共筛选出42个活性化合物,74个共有靶点,分析出91条生物过程,包括细胞因子受体结合、受体配体活性、受体调节活性、转录因子活性调节序列特异性DNA结合、核受体的活动、辅因子结合等。密切相关的通路有AGE-RAGE信号通路、TNF信号通路、IL-17信号通路、Toll样受体信号通路、人巨细胞病毒感染、C型凝集素受体信号通路、Th17细胞分化等;细胞实验表明三黄汤能够下调PI3K、AKT蛋白表达。结论:本研究揭示了三黄汤通过AGE-RAGE信号通路调控IL-1β、IL-6、IFN-γ等细胞因子发挥治疗湿疹的作用,为探讨三黄汤治疗湿疹的物质基础、作用机制等提供参考。

Objective:To predict the effective components and underlying mechanism of Sanhuang decoction in treating eczema based on network pharmacology and combined with the in vitro inflammation model of THP-1 macrophages.Method:The active chemical components in Sanhuang decoction were screened by TCMSP,and the common targets of compounds and eczema were searched through the GeneCards database.The“component-target-disease”network was constructed by using Cytoscape software,the protein-protein interaction(PPI)network was analyzed through the STRING database,and gene function annotation analysis(GO)and signaling pathway enrichment analysis(KEGG)were performed by the David database to predict the mechanism of Sanhuang decoction in treating eczema.THP-1 cells were stimulated with PMA and lipopolysaccharide(LPS)and intervened with Sanhuang decoction,the expression of inflammatory factors was detected by qRT-PCR and Elisa,and the expression of related proteins was detected by Western blot.Result:A total of 42 active compounds and 74 common targets were screened,and 91 biological processes were analyzed including cytokine receptor binding,receptor ligand activity,receptor regulatory activity,transcription factor activity regulation sequence specific DNA binding,nuclear receptor activity,cofactor binding,etc.The closely related pathways included AGE-RAGE,TNF,IL-17,and toll like receptor signaling pathway,human cytomegalovirus infection,C-type lectin receptor signaling pathway and Th17 cell differentiation.The cell experiments showed that Sanhuang decoction could downregulate the expression of PI3K and AKT proteins.Conclusion:This study reveals that Sanhuang decoction has effects of treating eczema by regulating cytokines such as IL-1β,IL-6 and IFN-γthrough the AGE-RAGE signaling pathway,which provides a reference for exploring the material basis and mechanism of Sanhuang decoction in treating eczema.