基于网络药理学分析连翘治疗牛乳房炎的作用机制及体内验证  

Mechanism of Forsythiae fructus in the treatment of mastitis based on network pharmacology and in vivo pharmacological evaluation

作　　者：王小榛 刘畅[1] 宋雪娇 邢晨 唐阳 张岳峰 吴桐 陈雪英[1] 李艳华[1] 刘艳艳[1] WANG Xiaozhen;LIU Chang;SONG Xuejiao;XING Chen;TANG Yang;ZHANG Yuefeng;WU Tong;CHEN Xueying;LI Yanhua;LIU Yanyan(College of Veterinary Medicine,Northeast Agricultural University,Harbin 150036,China)

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150036

出　　处：《中国兽医学报》2022年第12期2534-2542,共9页Chinese Journal of Veterinary Science

基　　金：黑龙江省重点研发计划资助项目(GA21B006-1)。

摘　　要：基于网络药理学和体内动物试验探究连翘(Forsythiae fructus,FF)治疗牛乳房炎的作用机制。利用TCMSP数据库检索连翘的成分和作用靶点,并与通过DisGeNET、GeneCards和OMIM数据库检索出的乳房炎的疾病靶点交集整合,获得连翘治疗乳房炎的潜在靶点。运用Cytoscape 3.6.1构建成分-靶点-疾病生物网络图,同时结合STRING平台构建PPI网络,通过DAVID平台及Cytoscape插件GlueGO进行GO和KEGG富集分析。构建金黄色葡萄球菌诱导的小鼠乳房炎模型,进行组织病理学、荧光定量PCR(qRT-PCR)及酶联免疫吸附(ELISA)分析。结果显示,连翘23个潜在成分对应208个靶点,牛乳房炎167个靶点,药物-疾病交集靶点25个。PPI核心靶点有白介素-6(interleukin-6,IL-6)、肿瘤坏死因子(tumor necrosis factor,TNF)等,主要参与免疫反应、细胞因子活性等过程,涉及IL-17信号通路、NF-κB信号通路、TNF信号通路等。体内试验结果表明,连翘能改善金黄色葡萄球菌诱导的小鼠乳房炎的炎症损伤。qRT-PCR及ELISA试验结果显示,连翘用药组IL-17/NF-κB信号通路关键基因表达量及蛋白含量均显著低于模型组(P<0.05或P<0.01),揭示连翘抗乳房炎作用与IL-17/NF-κB信号通路有关。从网络药理学角度初步揭示了连翘治疗乳房炎的多成分-多靶点-多通路的特点,为探究连翘治疗乳房炎作用机制提供了理论依据。This study aims to explore the mechanism of action of Forsythiae fructus(FF)in the treatment of bovine mastitis based on network pharmacology and in vivo animal testing.The components and targets of action of FF were retrieved using the TCMSP database and integrated with the intersection of disease targets of mastitis retrieved through the DisGeNET,GeneCards and OMIM databases to obtain potential targets of FF for the treatment of mastitis.Cytoscape 3.6.1 was used to construct the component-target-disease biological network map,and also combined with STRING platform to construct PPI network,GO and KEGG enrichment analysis was performed by DAVID platform and Cytoscape plug-in GlueGO.A mouse mastitis model induced by Staphylococcus aureus was constructed and analyzed by histopathology,qRT-PCR and ELISA.The results showed that 23 potential components of FF corresponded to 208 targets,167 targets for bovine mastitis,and 25 targets for drug-disease intersection.The core targets of PPI included interleukin-6(IL-6)and tumor necrosis factor(TNF),which were mainly involved in the immune response and cytokine activity,including IL-17 signaling pathway,NF-κB signaling pathway,TNF signaling pathway,etc.The results of in vivo experiments showed that FF could alleviate the inflammation of mammary glands in mice.The results of qRT-PCR and ELISA showed that the IL-17/NF-κB signaling pathway-related gene expression and protein content were significantly lower in the FFadministered group than in the model group(P<0.05 or P<0.01),which revealed that FF could play a role in the treatment of mastitis by regulating IL-17/NF-κB signaling pathway.This paper initially reveals the multicomponent-multitarget-multipathway characteristics of FF in the treatment of mastitis from the perspective of network pharmacology,which provides a theoretical basis for exploring the mechanism of action of FF in the treatment of mastitis.

关 键 词：连翘 乳房炎 网络药理学 炎症 IL-17/NF-κB信号通路 

分 类 号：S857.2[农业科学—临床兽医学] S859.7[农业科学—兽医学]