基于网络药理学的益气解毒复方对实验性自身免疫性重症肌无力大鼠的免疫治疗效果及机制  

Effect and mechanism of Yiqi Jiedu decoction based on network pharmacology on the immunotherapy of experimental autoimmune rats with myasthenia gravis

作　　者：沙婷 高霄 邓于新 彭亚倩 唐智 况时祥[3] 齐晓岚[1] SHA Ting;GAO Xiao;DENG Yuxin;PENG Yaqian;TANG Zhi;KUANG Shixiang;QI Xiaolan(Key Laboratory of Endemic and Minority Diseases of the Ministry of Education,Guizhou Medical University&Key Laboratory of Medical Molecular Biology of Guizhou,Guiyang 550004,Guizhou,China;Department of Laboratory,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Neurology,the Second Affiliated Hospital of Guiyang College of Traditional Chinese Medicine,Guiyang 550000,Guizhou,China)

机构地区：[1]贵州医科大学地方病与少数民族疾病教育部重点实验室&贵州省医学分子生物学重点实验室,贵州贵阳550004 [2]贵州医科大学附属医院检验科,贵州贵阳550004 [3]贵阳中医学院第二附属医院神经内科,贵州贵阳550000

出　　处：《贵州医科大学学报》2024年第10期1405-1416,1427,共13页Journal of Guizhou Medical University

基　　金：国家自然科学基金(82260263);贵州省科技计划项目(黔科合支撑〔2023〕一般232)。

摘　　要：目的基于网络药理学探讨益气解毒复方对实验性自身免疫性重症肌无力(MG)大鼠免疫治疗的效果及机制。方法运用中医药分子机理生物信息学分析软件(BATMAN-TCM)筛选益气解毒复方相关靶点信息,利用在线人类孟德尔遗传(OMIM)数据库以及Genecards数据库查询MG疾病有关靶点的信息;通过注释、可视化和综合发现数据库(DAVID)、Metascape数据库对益气解毒复方治疗MG的潜在靶点进行蛋白-蛋白互作网络(PPI)分析、基因本体(GO)功能富集及京都基因组百科全书(KEGG)相关信号通路分析;制备鼠源性乙酰胆碱受体(AChR)Rα97-116肽段抗原乳剂并皮下注射至30只无特定病原体(SPF)级雌性Lewis大鼠的尾基部、后肢垫部及背部(第1次免疫注射),分别于第30天和第45天时进行加强免疫(第2、3次免疫注射),第3次免疫注射结束后1周判断实验性自身免疫性MG(EAMG)大鼠模型是否成功,造模成功后EAMG大鼠随机均分为模型组、0.945 g/(kg·d)益气解毒复方(低剂量)组、1.890 g/(kg·d)益气解毒复方(中剂量)组、3.780 g/(kg·d)益气解毒复方(高剂量)组及5.400 mg/(kg·d)醋酸泼尼松片剂组,另取6只SPF级雌性Lewis大鼠作为正常组,各药物组大鼠分别予对应药物、正常组和模型组大鼠分别予等体积纯净水,连续灌胃干预30 d,采用电子天平和Lennon评分检测各组大鼠灌胃干预前和干预第30天时的体质量和肌力,同时采用酶联免疫吸附试剂盒检测各组大鼠血清AChR抗体(AChR-Ab)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)的水平,采用流式细胞术方法检测各组大鼠灌胃干预第30天时外周血中白细胞分化抗原4(CD4^(+))T细胞、白细胞分化抗原8(CD8^(+))T细胞的百分比及T淋巴细胞亚群CD4^(+)/CD8^(+)比率;灌胃干预30 d后处死各组大鼠,取骨骼肌组织,采用苏木精-伊红染色(HE)评估骨骼肌组织的形态学特征。结果筛选到益气解毒�Objective To explore the effect and mechanism of Yiqi Jiedu decoction on the immunotherapy of experimental autoimmune rats with myasthenia gravis(MG)based on network pharmacology.Methods Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMAN-TCM)was used to screen the target information related to Yiqi Jiedu decoction,and the Online Mendelian Inheritance in Man(OMIM)database and Genecards database was utilized to search for target information related to MG.The protein-protein interaction(PPI)network analysis,gene ontology(GO)functional enrichment,and Kyoto encyclopedia of genes and genomes(KEGG)pathway analysis were performed through annotation,visualization,and comprehensive discovery in the Database for Annotation,Visualization and Integrated Discovery(DAVID)and Metascape database for potential targets of Yiqi Jiedu decoction in the treatment of MG.The rat acetylcholine receptor(AChR)Rα97-116 peptide antigen emulsion was prepared and subcutaneously injected into the tail base,hind paw,and back of 30 specific pathogen-free(SPF)female Lewis rats(as the first immunization);and booster immunity was carried out on the 30 th day and 45 th day respectively(as the second and third immunizations).Whether the rat model of experimental autoimmune MG(EAMG)was successfully established was assessed one week after the third immunization.After successful modelling,EAMG rats were divided into the model group,Yiqi Jiedu decoction groups with doses of 0.945 g/(kg·d)as the low dose group,1.890 g/(kg·d)as the medium dose group and 3.780 g/(kg·d)as the high dose group,and the prednisone acetate tablet group with a dose of 5.400 mg/(kg·d),while other 6 SPF female Lewis rats were collected as the normal group.The drug groups were treated with corresponding drugs,and the normal and model groups were just given pure water,all intervened by gavage for 30 days.The body weight and muscle strength of each group were measured by the electronic balance and Lennon score,and tail vein blood was extracte

关 键 词：大鼠 重症肌无力 益气解毒复方 网络药理学 CD4^(+)/CD8^(+)比值 乙酰胆碱受体抗体 

分 类 号：R746.1[医药卫生—神经病学与精神病学]