摘要
该研究通过整合网络药理学和动物实验研究藏族药六锐胶囊干预实验性自身免疫性葡萄膜炎(experimental autoimmune uveitis, EAU)的作用机制,为深入探讨六锐胶囊治疗EAU的药效机制提供实验依据与参考。结合中医药百科全书数据库、BATMAN-TCM数据库与文献筛选六锐胶囊的活性成分与靶点;借助GEO、GeneCards、OMIM、TTD数据库检索EAU相关靶点,并将药物靶点与疾病靶点取交集;运用STRING数据库进行蛋白-蛋白互作(protein-protein interaction, PPI)网络分析;借助g:Profiler数据库进行GO(Gene Ontology)富集分析和KEGG(Kyoto Encyclopedia of Genes and Genomes)通路富集分析。此外,通过光感受器间维生素A类结合蛋白(interphotoreceptor retinoid-binding protein, IRBP)诱导塑造EAU大鼠模型,给予六锐胶囊进行干预,通过观察各组大鼠眼前节炎性反应、大鼠视网膜组织病理形态及采用实时荧光定量PCR(q-PCR)和蛋白质免疫印迹法(Western blot)检测六锐胶囊治疗EAU的作用机制。网络药理学分析预测了六锐胶囊与EAU的共同靶点51个;GO富集分析和KEGG分析结果主要涉及白细胞介素17(interleukin-17,IL-17)信号通路、肿瘤坏死因子(tumor necrosis factor, TNF)信号通路、核因子κB(nuclear factor-kappa B,NF-κB)信号通路、脂质体受体等生物学过程与通路。动物实验结果提示成功构建EAU大鼠模型;与空白组比较,模型组大鼠眼前节的炎性反应明显,眼组织DLL4(delta-like 4)、Notch1、IL-17、TNF-α蛋白与基因表达升高。与模型组比较,六锐胶囊能减轻大鼠眼前节与视网膜的炎性反应,降低眼组织DLL4、Notch1、IL-17、TNF-α蛋白与基因表达。藏族药六锐胶囊可降低眼组织DLL4/Notch1/IL-17信号通路相关基因与蛋白表达,减轻眼部炎性反应,可能是其治疗EAU的效用机制之一。
By integrating network pharmacology and animal experiments, we studied the pharmacodynamic mechanism of the Tibetan medicine Liurui Capsules in the treatment of experimental autoimmune uveitis(EAU). The active ingredients and targets of Liurui Capsules were searched against the Encyclopedia of Traditional Chinese Medicine(ETCM), Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMAN-TCM), and relevant literatures. The EAU-related targets were obtained from Gene Expression Omnibus(GEO), GeneCards, Online Mendelian Inheritance in Man(OMIM), and Therapeutic Target Database(TTD). The common targets shared by Liurui Capsules and EAU were identified, and the protein-protein interaction(PPI) network was established via STRING. Gene Ontology(GO) annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were conducted via g: Profiler. The rat model of EAU was induced by interphotoreceptor retinoid-binding protein(IRBP) and treated with Liurui Capsules. The inflammatory response of anterior segment and the pathological morphology of retina were observed. The mRNA and protein levels of delta-like ligand 4(DLL4), Notch1, interleukin-17(IL-17), and tumor necrosis factor-alpha(TNF-α) were determined by real-time quantitative PCR(q-PCR) and Western blot, respectively. The network pharmacology analysis predicted 51 common targets of Liurui Capsules and EAU, which were mainly involved in IL-17, TNF, and nuclear factor-kappa B(NF-κB) signaling pathways, as well as liposome receptors and other biological processes. Compared with the control group, the modeling of EAU caused inflammatory changes in the anterior segment and retina and up-regulated mRNA and protein levels of DLL4, Notch1, IL-17, and TNF-α in ocular tissue. Compared with the model group, Liurui Capsules reduced the inflammatory reaction of anterior segment and retina and down-regulated the mRNA and protein levels of DLL4, Notch1, IL-17, and TNF-α. Liurui Capsules can down-regulate the expression of the proteins involved in DLL4/Notch1/IL-17 signaling pathway in ocular tissue and alleviate the ocular inflammation, which may be one of the mechanisms of Liurui Capsules in the treatment of EAU.
作者
边猛
周政
朱春胜
聂安政
李晓萍
BIAN Meng;ZHOU Zheng;ZHU Chun-sheng;NIE An-zheng;LI Xiao-ping(the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2022年第24期6687-6695,共9页
China Journal of Chinese Materia Medica
基金
河南省医学科技攻关项目(LHGJ20200365)
郑州大学第一附属医院横向课题(K2020-0123)
关键词
六锐胶囊
实验性自身免疫性葡萄膜炎
网络药理学
药效机制
Liurui Capsules
experimental autoimmune uveitis
network pharmacology
pharmacodynamic mechanism
作者简介
边猛,主管药师,研究方向为临床中药学,E-mail:bianmeng0208@163.com;通信作者:李晓萍,主任药师,研究方向为医院药学管理,E-mail:lixiaoping630209@163.com
