网络药理学预测虎杖-吴茱萸抗痛风作用及验证研究

Prediction and Validation of Anti Gout Effect of Polygonum Cuspidatum andEvodia Rutaecarpa by Network Pharmacology

目的:通过网络药理学和实验验证方法探讨虎杖-吴茱萸配伍抗痛风(gouty arthritis,GA)的作用机制。方法:运用中药系统药理学数据库(traditional chinese medicine systems pharmacology database and analysis platform,TCMSP)筛选出虎杖和吴茱萸的活性成分及相关靶点蛋白,采用Cytoscape v3.7.2软件构建活性成分-靶点网络图,借助STRING数据库构建蛋白相互作用网络(protein-protein interaction network,PPI),再利用DAVID数据库进行基因本体(gene ontology,GO)功能与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路富集分析,预测虎杖-吴茱萸配伍抗痛风的作用机制,进而建立大鼠急性GA模型,采用比色法测定大鼠血清中尿酸(uric acid,UA)与黄嘌呤氧化酶(xanthine oxidase,XOD)水平,并采用ELISA试剂盒检测大鼠血清中白介素-6(interleukin 6,IL-6)、白介素-1β(interleukin 1β,IL-1β)、白介素-10(interleukin 10,IL-10)及白介素-17β(interleukin 17β,IL-17β)指标的含量变化。结果:从虎杖-吴茱萸中共筛选得到26种有效成分,与痛风相关的靶点有21个,其中炎症因子发挥重要作用。验证实验也表明,虎杖-吴茱萸各剂量组能显著降低血清中UA和XOD水平,能显著降低IL-6、IL-1β、IL-17β的含量并增加IL-10的水平。结论:虎杖-吴茱萸的活性成分可调控GA发病重要通路中的多个靶点,初步实验验证其抗GA的作用机制可能是通过炎症相关通路,调控炎症因子发挥作用,为其临床应用提供参考。

Objective:To predict and validate the active compounds and mechanism of Polygonum Cuspidatum and Evodia Rutaecarpa against gout arthritis(GA)by network pharmacology.Methods:The active components and related target proteins of Polygonum Cuspidatum and Evodia Rutaecarpa were obtained by searching Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)database.Cytoscape software(version v3.7.2)was used to build a network diagram between active components and targets.Targets retrieved were used to build a protein-protein interaction network(PPI)network based on the analysis from STRING database.Gene ontology(Go)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis was then conducted by the DAVID database to elucidate the pathway involved in the Polygonum Cuspidatum and Evodia Rutaecarpa against GA.Then,A rat model of GA was used to further investigate the mechanism,the levels of uric acid(UA),xanthine oxidase(XOD),interleukin 6(IL-6),interleukin 1β(IL-1β),interleukin 10(IL-10)and interleukin 17β(IL-17β)in rat serum were measured by colorimetry and ELISAs.Results:There were a total of 26 active components and 21 targets of Polygonum Cuspidatum and Evodia Rutaecarpa against GA.Inflammatory factors play an important role.Experimental validation suggests that Polygonum Cuspidatum and Evodia Rutaecarpa significantly decreased the contents of UA,XOD,IL-6,IL-1βand IL-17β,and increased the level of IL-10 in serum.Conclusion:The active components of Polygonum Cuspidatum and Evodia Rutaecarpa can regulate multiple targets in the important pathway of GA.Animal experiments preliminarily verify that the mechanism of Polygonum Cuspidatum and Evodia Rutaecarpa may be through inflammation related pathways,regulating inflammatory factors to treat GA,which provides evidence to support its clinical use.