白虎加人参汤治疗肥胖症合并2型糖尿病的潜在活性成分及作用机制分析  

Analysis of Potential Active Ingredients and Mechanism of Baihu Jia Renshentang in Treatment of Obesity Complicated with Type 2 Diabetes Mellitus

作　　者：谭丹妮 向琴 俞赟丰 喻嵘 TAN Danni;XIANG Qin;YU Yunfeng;YU Rong(School of Traditional Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学中医学院,长沙410208

出　　处：《中国实验方剂学杂志》2024年第13期1-10,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金联合基金重点支持项目(U21A20411);国家自然科学基金面上项目(82074400);湖南省重点研发计划项目(2020SK2101);湖南省教育厅创新平台开放基金项目(20K094)。

摘　　要：目的:通过网络药理学结合体内实验验证,探讨白虎加人参汤治疗肥胖症合并2型糖尿病(T2DM)的潜在活性成分和作用靶点。方法:利用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法(UPLC-Q-Exactive Orbitrap MS)分析鉴定白虎加人参汤物质基础。在ChEMBL、疗效药靶数据库(TTD)、YaTCM、DisGeNET和中药免疫肿瘤学(TCMIO)等数据库中查询活性成分作用的潜在靶点,并将这些靶点与疾病靶点取交集确定共同靶点。将共同靶点导入STRING数据库构建蛋白质-蛋白质相互作用(PPI)网络,通过cytoHubba插件确定hub基因,利用分子对接验证hub基因与白虎加人参汤中生物活性成分的结合能。同时,将共同靶点导入DAVID平台进行基因本体(GO)功能注释及京都基因和基因组百科全书(KEGG)通路富集分析。通过动物实验验证预测结果,将18只8周龄雄性骨骼肌胰岛素样生长因子-1受体功能缺失(MKR)小鼠高脂饮食饲养12周,制备肥胖症合并T2DM小鼠模型。随机将小鼠均分为模型组、二甲双胍组(0.2 g·kg^(-1))、白虎加人参汤组(以生药计27 g·kg^(-1)),另设6只同龄雄性FVB小鼠作为正常组,各组小鼠给予相应药物灌胃,正常组及模型组灌胃等量蒸馏水,1次/d,连续6周。给药结束后,检测各组小鼠的体质量、Lee's指数、空腹血糖(FBG)、口服糖耐量试验(OGTT),观察附睾白色脂肪组织病理形态,并采用实时荧光定量聚合酶链式反应(Real-time PCR)检测附睾白色脂肪组织中hub基因mRNA的表达水平。结果:共鉴定出白虎加人参汤200种化合物,其中64种生物活性成分反向匹配到384个靶点,与肥胖症合并T2DM相关的靶点共308个。hub基因包括丝裂原活化蛋白激酶1(MAPK1)、信号转导与转录激活因子3(STAT3)、MAPK3、白细胞介素(IL)-2、Janus酪氨酸蛋白激酶1(JAK1)、核转录因子-κB p65(RELA)、雌激素受体1(ESR1)、转录因子AP-1(JUN)、MAPK14和淋巴细胞特异性蛋白酪氨Objective:To investigate the potential active ingredients and targets of Baihu Jia Renshentang(BHJRST)for the treatment of obesity combined with type 2 diabetes mellitus(T2DM)by network pharmacology and in vivo experiments.Method:Ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS)was used to analyze and identify the material basis of BHJRST.Subsequently,potential targets for the action of the active ingredients were queried in databases such as ChEMBL,Therapeutic Target Database(TTD),YaTCM,DisGeNET and Traditional Chinese Medicine on Immuno-Oncology(TCMIO),and the shared targets were identified by taking the intersection of these targets with disease targets.The shared targets were imported into the STRING database to construct a protein-protein interaction(PPI)network,the hub genes were identified by cytoHubba plug-in,and molecular docking was used to validate the binding energy of the hub genes to the bioactive ingredients in BHJRST.Meanwhile,the shared targets were imported into the DAVID platform for gene ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.The predicted results were subsequently verified by animal experiments.Eighteen 8-weekold male skeletal muscle insulin-like growth factor-1 receptor dysfunction(MKR)mice were induced by a highfat diet for 12 weeks in order to prepare a mouse model of obesity combined with T2DM.The mice were randomly divided into the model group,metformin group(0.2 g·kg^(-1))and BHJRST group(27 g·kg^(-1) in raw material),and another 6 male FVB mice of the same age as the normal group.The mice in each group were were given the corresponding drugs by gavage,and the normal and model groups were given the same amount of distilled water by gavage,1 time/d for 6 consecutive weeks.At the end of administration,the body mass,Lee's index,fasting blood glucose(FBG),oral glucose tolerance tes(t OGTT)of mice in each group were examined,an

关 键 词：白虎加人参汤 2型糖尿病 肥胖 胰岛素抵抗 网络药理学 超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法(UPLC-Q-Exactive Orbitrap MS) hub基因 

分 类 号：R22[医药卫生—中医基础理论] R587.1[医药卫生—中医学] R28[理学—分析化学] O657[理学—化学]