消瘀止痛合剂治疗膝骨关节炎作用机制的网络药理学与动物实验验证  

Network Pharmacology and Animal Experimental Validation of the Mechanism of Xiaoyu Zhitong Mixture in Treating Knee Osteoarthritis

作　　者：陈炜坚 韦志豪 郭诗雯 钟健[1,2] 石杰 郭俊 胡耶芳 唐刚健[2] CHEN Weijian;WEI Zhihao;GUO Shiwen;ZHONG Jian;SHI Jie;CUO Jun;HU Yefang;TANG Gangjian(Guangxi University of Chinese Medicine,Nanning 530000 Guangxi,China;Guilin Municipal Hospital of Traditional Chinese Medicine,Guilin 541002 Guangxi,China)

机构地区：[1]广西中医药大学,广西南宁530000 [2]桂林市中医医院,广西桂林541002

出　　处：《中药新药与临床药理》2023年第8期1095-1104,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：吴阶平医学基金会临床科研专项资助基金(320.6750.2022-20-32);广西中医药大学研究生教育创新计划项目(YCSY2022052);桂林市科学研究与技术开发计划项目(2020011208-1)。

摘　　要：目的通过网络药理学方法预测消瘀止痛合剂(桃红四物汤合玄胡索散化裁)治疗膝骨关节炎(knee osteoarthritis,KOA)的活性成分、作用靶点与信号通路,通过动物实验验证其作用机制。方法消瘀止痛合剂的总活性成分及靶点通过中药系统药理学数据库与分析平台、成分靶点预测数据库和文献补充等途径获取;通过在线人类孟德尔遗传病数据库(OMIM)、GeneCards、PharmGkB数据库获取膝骨关节炎疾病靶点;使用Venn平台绘制文氏图,利用STRING数据库及CytoScape软件建立基于活性成分与疾病共同靶点的蛋白相互作用(PPI)网络;利用Metascape平台进行基因本体(GO)富集分析与京都基因与基因组百科全书(KEGG)通路分析。动物实验通过木瓜蛋白酶诱导大鼠膝骨关节炎模型进行实验验证。结果获得消瘀止痛合剂280种活性成分,共同作用靶点172个,涉及肿瘤坏死因子(TNF)、白细胞介素(IL)-1β、胱天蛋白酶(Caspase)-3等治疗核心靶点,参与对无机物的反应、细胞迁移的正向调控、对脂多糖的反应等7259个生物过程;涉及IL-17、TNF、丝裂原活化蛋白激酶、磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)等261条信号通路。动物实验结果中,苏木精-伊红染色、番红O-固绿染色、Mankin评分表明消瘀止痛合剂对膝骨关节炎软骨修复疗效明显(P<0.05)。消瘀止痛合剂通过下调IL-1β、TNF-α、Caspase-3蛋白表达从而治疗膝骨关节炎(P<0.05),验证了网络药理学的部分预测。结论消瘀止痛合剂可通过多靶点、多通路治疗膝骨关节炎。证实其通过下调IL-1β、TNF-α、Caspase-3蛋白表达,从而减缓软骨退变、促进软骨修复,达到治疗膝骨关节炎的效果。该研究为临床使用及后续研究提供了实验依据。Objective To preliminarily predict the active components,targets,and signal pathways of Xiaoyu Zhitong Mixture(Taohong Siwu Decoction combined with Xuanhusuo Powder)in the treatment of knee osteoarthritis(KOA)through network pharmacological methods,and to verify the mechanism of action through animal experiments.Methods The total active components and targets of Xiaoyu Zhitong Mixture were obtained through Traditional Chinese Medicine Database and Analysis Platform,ingredient target prediction database and literature supplement.The KOA disease targets were obtained through Online Mendelian Inheritance in Man(OMIM),GeneCards,and PharmGkB databases.We used Venn platform to draw Venn diagram.STRING database and CYTOSCAPE software were adopted to establish a protein-protein interaction(PPI)network based on the common target of active components and diseases.The Metascape platform was used to perform gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis.Rat model with papain-induced KOA was used to verify the simulated results.Results A total of 280 active components of Xiaoyu Zhitong Mixture and 172 common targets were obtained,involving the core therapeutic targets such as tumor necrosis factor(TNF),interleukin(IL)-1β,and Caspase-3;7259biological processes were found,including response to inorganic substance,positive regulation of cell migration and response to lipopolysaccharide.A total of 261 signal pathways were obtained,including IL-17,TNF,mitogenactivated protein kinase(MAPK),phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt).The hematoxylineosin staining,safranine O-fast green staining,and Mankin score in animal experiments showed that Xiaoyu Zhitong Mixture had a significant effect on cartilage repair in KOA(P<0.05).Xiaoyu Zhitong Mixture can down-regulate protein expressions of IL-1β,TNF-α,and Caspase-3 for the treatment of KOA(P<0.05).These results confirmed partial predictions of network pharmacology.Conclusion Xiaoyu Zhitong Mixture can treat KOA

关 键 词：膝骨关节炎 消瘀止痛合剂 桃红四物汤 玄胡索散 IL-1Β TNF-α Caspase-3 网络药理学 作用机制 大鼠 

分 类 号：R285.5[医药卫生—中药学]