基于网络药理学及实验验证探讨山芋丸对“痹症”模型大鼠的治疗作用及其机制  

Therapeutic Effect and Mechanism of Shanyu Pills in Treating"Arthromyodynia"Model Rats Based on Network Pharmacology and Experiment

作　　者：吴倩媛 刘翔翔 王英豪[1] 王颖峥 刘志臻[1,2] WU Qianyuan;LIU Xiangxiang;WANG Yinghao;WANG Yingzheng;LIU Zhizhen(Fujian University of Traditional Chinese Medicine,College of Pharmacy,Fuzhou 350122,China;Fujian University of Traditional Chinese Medicine,Science and Technology Branch,Fuzhou 350122,China)

机构地区：[1]福建中医药大学药学院,福州350122 [2]福建中医药大学科学技术处,福州350122

出　　处：《中国现代应用药学》2024年第19期2623-2634,共12页Chinese Journal of Modern Applied Pharmacy

基　　金：国家中医药管理局临床中药学高水平中医药重点学科建设项目(国中医药人教函[2023]85号)。

摘　　要：目的通过网络药理学和动物实验初步探究山芋丸对佐剂大鼠踝关节痹症的治疗作用及机制,并从中医“痹症”与“扶正祛邪”的角度探讨其理论基础。方法通过TCMSP数据库对山药、白术、人参活性成分进行筛选与靶点预测。在GeneCards、OMIM、DrugBank中进行类风湿关节炎(rheumatoid arthritis,RA)靶点的检索及筛选。利用Cytoscape 3.9.1构建“药物-活性成分-疾病”网络图。将交集靶点输入到STRING数据库分析获得蛋白互作网络,导入Cytoscape 3.9.1软件进行可视化处理。使用Metascape对交集靶点进行GO及KEGG富集分析。通过AutoDock软件进行分子对接,并用Pymol进行结果可视化。采用弗氏完全佐剂复制佐剂性关节炎大鼠模型,将36只SD大鼠随机分为正常对照组、模型组、阳性药组(甲氨蝶呤灌胃给药)和山芋丸低(4.61 g·kg^(-1))、中(9.22 g·kg^(-1))、高(18.43 g·kg^(-1))剂量组。测量大鼠的踝关节直径,计算大鼠的踝关节肿胀度;取大鼠的踝关节进行组织病理学观察,使用TUNEL细胞凋亡试剂盒检测大鼠滑膜细胞的凋亡程度,并测定血清中IL-6、TNF-α、MDA的含量,取大鼠脾脏并计算脾脏指数,采用qRT-PCR验证滑膜组织中MAPK3、Caspase 3 mRNA的表达,蛋白免疫印迹法检测踝关节组织中MAPK3表达水平。结果筛选出54个有效成分,对应于山芋丸的325个靶标。此外,845个基因与RA密切相关,其中有86个与山芋丸的靶点重叠。功能富集分析表明,山芋丸通过调节多种途径发挥其在RA中的治疗作用。给药组肿胀度相较于模型组明显降低(P<0.05)。HE染色结果显示,山芋丸能抑制滑膜肿胀程度。TUNEL细胞凋亡结果显示,山芋丸给药后能够抑制滑膜细胞凋亡。与模型组相比,给药组的IL-6、TNF-α、MDA水平以及脾脏指数均降低(P<0.05);大鼠踝关节组织中的MAPK3、Caspase 3 mRNA表达和MAPK3蛋白表达也明显降低(P<0.05)。结论山芋丸能减轻佐剂模型�OBJECTIVE To investigate the therapeutic effects of Shanyu pills on ankle arthrogryposis in adjuvant rats through network pharmacology and animal experiments and to explore the theoretical basis of using Shanyu pills to treat rheumatoid arthritis(RA)from the perspective of"arthromyodynia"and"strengthening vital Qi to eliminate pathogenic factor"in traditional Chinese medicine.METHODS The active ingredients of Dioscoreae Rhizoma,Atractylodis Macrocephalae Rhizoma,and Ginseng Radix et Rhizoma were screened and target predicted using the TCMSP database.RA targets were searched and screened in GeneCards,OMIM,and DrugBank.A"drug-active ingredient-disease"network diagram was constructed using Cytoscape 3.9.1.The intersection targets were input into the STRING database for analysis to obtain protein interaction networks and then imported into Cytoscape 3.9.1 for visualization.Metascape was used to perform GO and KEGG enrichment analyses on intersecting targets,and then a microbiome platform was used for enrichment analysis and visualization.In addition,molecular docking was performed using AutoDock,and the results were visualized using Pymol.A rat model of adjuvant arthritis was established using Freund’s complete adjuvant.Thirty-six SD rats were randomly divided into the normal control group,the model group,the positive drug group,the low-dose group of Shanyu pills(4.61 g·kg−1),the medium-dose group of Shanyu pills(9.22 g·kg−1),and the high-dose group of Shanyu pills(18.43 g·kg−1).The diameter of the ankle joint of rats was measured,and the swelling degree of the ankle joint was calculated.The ankle joint of rats was removed for histopathological observation,and the TUNEL reagent was used to detect the degree of the apoptosis of synovial cells in rats and to determine the level of IL-6 and TNF-α.Moreover,the content of MDA was measured by taking the spleen and calculating the spleen index.The expression level of MAPK3 and Caspase 3 mRNA in synovial tissue was detected using qRT-PCR.RESULTS A total of 54 eff

关 键 词：山芋丸 痹症 类风湿关节炎 网络药理学 实验验证 

分 类 号：R285.5[医药卫生—中药学]