抑肝散治疗阿尔茨海默病的作用及机制探讨  

Treatment and mechanism of Yigansan on Alzheimer's disease

作　　者：张然[1] 焦扬[2] 邹爱英[3] 李睿 张秋阳[5] 商倩 ZHANG Ran;JIAO Yang;ZOU Aiying;LI Rui;ZHANG Qiuyang;Shang Qian(School of Pharmacy and Biotechnology,Tianjin Medical College,Tianjin 300222,China;Department of Pharmacy,First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300193,China;Department of Pharmacy,The Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300250,China;School of Chemical Engineering,Tianjin University,Tianjin 300073,China;Department of Basic Medicine,Tianjin Haihe Hospital,Tianjin 300350,China;Tianjin Key Laboratory of Molecular Design and Drug Discovery,Tianjin Institute of Pharmaceutical Research,Tianjin300462,China)

机构地区：[1]天津医学高等专科学校药学与生物技术学院,天津300222 [2]天津中医药大学第一附属医院药学部,国家中医针灸临床医学研究中心,天津300193 [3]天津中医药大学第二附属医院药学部,天津300250 [4]天津大学化工学院,天津300073 [5]天津市海河医院基础医学实验部,天津300350 [6]天津药物研究院有限公司天津市新药设计与发现重点实验室,天津300462

出　　处：《药物评价研究》2024年第8期1749-1759,共11页Drug Evaluation Research

基　　金：天津市中医药重点领域科研项目(2022008)。

摘　　要：目的探究抑肝散对阿尔茨海默病(AD)的治疗作用,通过网络药理学方法预测其机制,并进行实验验证。方法以D-半乳糖诱导AD小鼠模型,通过水迷宫和八臂迷宫实验考察抑肝散(400、800 mg·kg^(-1))对AD小鼠学习记忆功能的影响,并通过实时荧光定量PCR(qRT-PCR)法分析AD小鼠脑组织中肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)、白细胞介素-2(IL-2)、白细胞介素-6(IL-6)的mRNA表达变化。通过中药系统药理学数据分析平台(TCMSP)对抑肝散的组方药材柴胡、甘草、川芎、当归、白术、茯苓和钩藤进行活性成分筛选,采用GeneCards等数据库获取AD疾病靶点,筛选其核心靶点,并对核心靶点进行基因本体(GO)和京都基因与基因组百科全书(KEGG)通路富集分析,分别获取核心靶点涉及的生物过程(BP)、分子功能(MF)和细胞组分(CC)以及KEGG信号通路。小鼠神经母细胞瘤Neuro-2a细胞和RAW264.7巨噬细胞分别与H_(2)O_(2)和抑肝散(3、6μg·mL^(-1))共培养,观察抑肝散对Neuro-2a细胞体外增殖和凋亡的影响,及对Neuro-2a细胞Caspase-3和Caspase-8活性、活性氧(ROS)、谷胱甘肽(GSH)、丙二醛(MDA)、超氧化物歧化酶(SOD)和线粒体膜电位(MMP)的影响;观察抑肝散对RAW264.7细胞TNF-α和IL-1β、IL-2、IL-6表达的影响,对网络药理学结果进行验证。结果抑肝散可缓解AD小鼠学习记忆功能损伤,通过网络药理学技术筛选得到抑肝散治疗AD的活性成分159个,度值前10位的化合物分别是槲皮素、山柰酚、异鼠李素、β-谷甾醇、豆甾醇、7-甲氧基异黄酮、芒柄花素、柚皮素、美迪紫檀素和甘草查尔酮A,筛选得到核心靶点227个,度值排名前10的核心靶点为丝氨酸/苏氨酸蛋白激酶1(AKT1)、肿瘤坏死因子(TNF)、白细胞介素6(IL6)、肿瘤蛋白P53(TP53)、白细胞介素1β(IL1B)、雌激素受体1(ESR1)、原癌基因(JUN)、前列腺素氧化环化酶2(PTGS2)、半胱氨酸蛋白酶3(CASP3)和信�Objective To explore the therapeutic effect of Yigansan on Alzheimer's disease(AD),to predict its mechanism through network pharmacology,and to carry out experimental verification.Methods The effects of Yigansan(400 and 800 mg·kg-1)on the learning and memory function of AD mice were investigated in a D-galactose-induced AD mouse model,and the mRNA expressions of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-2(IL-2)and interleukin-6(IL-6)in the brain tissues of AD mice were analyzed by real-time quantitative PCR(qRT-PCR).Through the TCM Systematic Pharmacology Data Analysis Platform(TCMSP),the active ingredients of the herbal medicines Bupleuri Radix,Glycyrrhizae Radix et Rhizoma,Chuanxiong Rhizoma,Angelicae Sinensis Radix,Atractylodis Macrocephalae Rhizoma,Poria and Uncariae Ramulus cum Uncis were screened,and the AD targets were obtained by GeneCards and other databases,and the core targets were enriched by gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways.The biological processes(BP),molecular function(MF)and cellular composition(CC)and KEGG signaling pathwayinvolved in the core targets were obtained,respectively.Mouse neuroblastoma Neuro-2a cells and RAW264.7 macrophages were co-cultured with H_(2)O_(2)and hepatosuppress san(3 and 6μg·mL^(-1)),respectively,to observe the effects of Yigansan on the proliferation and apoptosis of Neuro-2a cells in vitro,and the activity of Caspase-3 and Caspase-8,reactive oxygen species(ROS),glutathione(GSH),malondialdehyde(MDA),effects on superoxide dismutase(SOD)and mitochondrial membrane potential(MMP).The effects of Yigansan on the expression of TNF-α,IL-1β,IL-2 and IL-6 in RAW264.7 cells were observed,and the network pharmacology results were verified.Results A total of 159 active ingredients were screened for the treatment of AD by network pharmacology technology,and the top 10 compounds were quercetin,kaempferol,isorhamnetin,β-sitosterol,stigmasterol,7-methoxyisoflavone,mangopertin,naringenin,meditalpin,and licochalcon

关 键 词：抑肝散 阿尔茨海默病 网络药理学 氧化损伤 炎症因子 

分 类 号：R285.5[医药卫生—中药学]