基于网络药理学、分子对接技术与实验验证探讨定志小丸干预阿尔兹海默病的机制研究  

Mechanism of Dingzhi Xiaowan(定志小丸)against Alzheimer's Disease:Based on Network Pharmacology,Molecular Docking,and Experimental Verification

作　　者：钟晓琴 宁振求 王凯[1] 蔡粤芳 邓敏贞[1,2] ZHONG Xiaoqin;NING Zhenqiu;WANG Kai;CAI Yuefang;DENG Minzhen(Guangzhou University of Chinese Medicine,Guangzhou 510006;The Second Affiliated Hospital of Guangzhou University of Chinese Medicine/Guangdong Provincial Hospital of Chinese Medicine/Guangdong Provincial Academy of Chinese Medical Sciences,Guangzhou 510120)

机构地区：[1]广州中医药大学,广州510006 [2]广州中医药大学第二附属医院/广东省中医院/广东省中医药科学院,广州510120

出　　处：《中药药理与临床》2023年第7期2-9,共8页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金资助项目(编号:81904104);广东省中医药局科研基金项目(编号:20211203);广东省中医院朝阳人才科研专项资助(编号:ZY2022KY06);中国博士后科研基金面上项目(编号:2021M690759)。

摘　　要：目的:应用网络药理学联合分子对接技术探讨定志小丸干预阿尔兹海默病(Alzheimer’s disease,AD)的作用机制。方法:通过TCMSP数据库及文献检索收集与查询定志小丸的组分化学成分信息;应用PubChem和SwissTargetPrediction数据库收集药物化学成分靶点、GeneCards数据库收集AD靶点;利用DAVID数据库对共有靶点进行基因本体(GO)功能及京都基因与基因组百科全书(KEGG)通路富集分析;使用Cytoscape软件构建成分与靶点等相关网络图,以筛选主要成分与靶点进行分子对接研究。采用水迷宫实验、ELISA和Western blot法检测定志小丸干预APP/PS1小鼠的作用情况。结果:共整理得到定志小丸的52个成分及其对应的881个靶点,AD靶点9599个,二者共同靶点101个,富集分析发现共同靶点主要参与细胞活力、缺氧应答等生物过程,及PI3K/AKT信号通路、缺氧诱导因子-1(HIF-1)信号通路、肿瘤坏死因子(TNF)信号通路等。分子对接结果显示,人参的二十碳-11,14,17-三烯酸甲酯、石菖蒲的环阿屯醇、茯苓的麦角甾烷-7,22-二烯-3β-醇及远志的3,6’-二芥子酰基蔗糖等成分能与热激蛋白90家族α类别A成员1(Heat Shock Protein 90 Alpha Family Class A Member 1,HSP90AA1)、表皮生长因子受体(Epidermal Growth Factor Receptor,EGFR)、Janus激酶3(Janus kinase 3,JAK3)、丝裂原活化蛋白激酶8(Mitogen-Activated Protein Kinase 8,MAPK8)等靶点良好对接。试验结果发现定志小丸明显改善APP/PS1小鼠的学习记忆能力,降低4-羟基壬烯酸(4-HNE)、白介素-6(IL-6)、TNF-α含量,下调磷酸化Janus激酶3(p-JAK3)和HSP90的表达,明显增加超氧化物歧化酶(SOD)活力和上调磷脂酰肌醇-4,5-二磷酸3-激酶催化亚单位α(PIK3CA)蛋白表达(P<0.05)。结论:定志小丸可能是通过二十碳-11,14,17-三烯酸甲酯、环阿屯醇、麦角甾烷-7,22-二烯-3β-醇和3,6’-二芥子酰基蔗糖等关键成分作用于HSP90、JAK3、PI3KCA等靶点改善神经元Objective:To explore the mechanism of Dingzhi Xiaowan(定志小丸)against Alzheimer's disease(AD)based on network pharmacology and molecular docking.Methods:The active components of Dingzhi Xiaowan were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and previous research,and targets of the components from PubChem and SwissTargetPrediction.AD-related targets were searched from GeneCards.DAVID was employed to identify the Gene Ontology(GO)terms and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways of the common targets,and Cytoscape to construct the networks on the components and targets to screen the main components and targets for molecular docking.Morris water maze,enzyme-linked immunosorbent assay(ELISA),and Western blotting were performed to examine the effect of Dingzhi Xiaowan on APP/PS1 mice.Results:A total of 52 components of Dingzhi Xiaowan,881 targets of the components,9599 AD-related targets,and 101 common targets were screened out.The enrichment suggested that the common targets were involved the major biological processes of cell viability and hypoxia response and the signaling pathways of phosphatidylinositol-3-kinase(PI3K)-protein kinase B(Akt)signaling pathway,hypoxia-inducible factor 1(HIF-1)signaling pathway,and tumor necrosis factor(TNF)signaling pathway.The results of molecular docking showed that methyl 11,14,17-eicosatrienoate of Renshen(人参),cycloartenol of Shichangpu(石菖蒲),ergostane-7,22-dien-3β-ol of Fuling(茯苓),and 3,6'-diespinoyl sucrose of Yuanzhi(远志)had high binding affinity with heat shock protein 90 kDa alpha A1(HSP90aA1),epidermal growth factor receptor(EGFR),Janus kinase 3(JAK3),and mitogen-activated protein kinase 8(MAPK8).The experiment indicated Dingzhi Xiaowan(定志小丸)significantly improved the learning and memory ability of APP/PS1 mice,decreased the levels of 4-hydroxynonenal(4-HNE),interleukin-6(IL-6),TNFα,phosphorylated(p)-JAK3,and HSP90,and increased the expression of superoxide dismutase(SOD

关 键 词：定志小丸 阿尔兹海默病 网络药理学 分子对接 

分 类 号：R285[医药卫生—中药学]