摘要
运用网络药理学方法探讨资木瓜治疗类风湿关节炎(rheumatoid arthritis,RA)的物质基础和作用机制,并通过分子对接和实验验证资木瓜抗RA的潜在靶点。从中药系统药理学数据库与分析平台筛选资木瓜中的活性成分及靶点。利用GeneCards、DisGeNET和OMIM数据库获取RA相关靶点,两者取交集,即资木瓜治疗RA的潜在作用靶点。利用Cytoscape 3.9.0软件,构建“中药-活性成分-共同靶点-疾病”网络。利用STRING建立蛋白相互作用(PPI)网络,运用RStudio 4.1.0软件对核心基因进行可视化。运用DAVID数据库进行基因本体(Gene Ontology,GO)功能注释和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析,预测其富集的信号通路并进行可视化展示。将筛选出来的活性成分作为配体,与筛选得到的RA核心靶点进行分子对接分析,最后通过动物实验进一步验证实验结果。资木瓜的主要活性成分4种、对应靶点137个,药物疾病共同靶点37个;GO富集分析得到239个条目(P<0.05),KEGG通路富集分析筛选出94条信号通路(P<0.05),主要涉及IL-17信号通路、肿瘤坏死因子信号通路、Toll样受体信号通路和NF-κB信号通路等。分子对接结果显示,资木瓜主要活性成分与RA核心靶点结合较好。动物实验结果显示,资木瓜可以缓解RA小鼠的关节肿胀度,显著降低白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)水平,显著降低血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)蛋白表达水平。资木瓜可通过影响VEGFA、IL-1β和IL6等核心靶点发挥抗炎和减少血管翳生成,从而减轻炎症,减少血管翳对关节的破坏以达到治疗RA的作用,以上研究为该传统中药日后治疗RA的进一步探索提供了新思路。
The material basis and mechanism of Chaenomelis Fructus in the treatment of rheumatoid arthritis(RA)were explored by network pharmacology,and the potential anti-RA targets of Chaenomelis Fructus were verified by molecular docking and animal experiments.The active components and targets of Chaenomelis Fructus were searched against the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform.GeneCards,DisGeNET,and OMIM were used to obtain RA-related targets.The common targets shared by Chaenomelis Fructus and RA were considered as the potential targets of Chaenomelis Fructus in the treatment of RA.Cytoscape 3.9.0 was employed to establish a“TCM-active component-common target-disease”network.The protein-protein interaction(PPI)network was established by STRING,and the core genes were visualized by RStudio 1.41717.0.DAVID was used for gene ontology(GO)annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment to predict and visualize the involved signaling pathways.Molecular docking was carried out with the active components screened out as ligands and RA core genes as the targets.Finally,the prediction results were verified by animal experiments.Four main active components of Chaenomelis Fructus were obtained,which corresponded to 137 targets.Chaenomelis Fructus and RA shared 37 c ommon targets.GO annotation yielded 239 terms(P<0.05),and KEGG pathway enrichment analysis screened out 94 signaling pathways(P<0.05),mainly involving interleukin-17(IL-17),tumor necrosis factor,Toll-like receptor,and nuclear factor-kappa B(NF-κB)signaling pathways.Molecular docking results showed that the main active components of Chaenomelis Fructus bound well with the core targets of RA.The results of animal experiments proved that Chaenomelis Fructus can alleviate joint swelling in the mic e with RA.The results of ELISA showed that Chaenomelis Fructus lowered the levels of interleukin-6(IL-6)and interleukin-1β(IL-1β).Western blot showed that Chaenomelis Fructus down-regulated the protein level of vascular endothelial growth factor A(VEGFA).Chaenomelis Fructus exerts anti-inflammatory effect and reduces pannus formation by regulating the core targets such as VEGFA,IL-1β,and IL6 in the treatment of RA.The findings of this study provide new ideas for the future treatment of RA with Chaenomelis Fructus.
作者
段志豪
金璨
邓颖
柳金浪
王颉
李世刚
周游
DUAN Zhi-hao;JIN Can;DENG Ying;LIU Jin-lang;WANG Jie;LI Shi-gang;ZHOU You(Department of Orthopedics,Affiliated Renhe Hospital of China Three Gorges University,Yichang 443001,China;Sports Medicine Research Institute of China Three Gorges University,Yichang 443001,China;Clinical Medical Research Center of Yichang Sports Injury and Repair,Yichang 443001,China;Medical College,China Three Gorges University,Yichang 443000,China;Three-Level Laboratory for Traditional Chinese Pharmacologic(Tumor)Research,Medical College,China Three Gorges University,Yichang 443000,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2023年第18期4852-4863,共12页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(82274170,81673665,81274166)
湖北省卫生健康委员会中医药科研项目(ZY2021M074)
湖北省自然科学基金项目(2021CFB414)。
关键词
网络药理学
分子对接
资木瓜
实验验证
作用机制
network pharmacology
molecular docking
Chaenomelis Fructus
experimental verification
mechanism of action
