摘要
目的运用网络药理学及分子对接的方法探究葛根素(Pue)干预阿尔茨海默病(AD)的分子作用机制,并进行动物实验验证。方法通过中药系统药理学数据库与分析平台(TCMSP)、集成药效团匹配平台(PharmMapper)等获取Pue的作用靶点,利用人类基因综合数据库(GeneCards)获取AD相关靶点,映射后获取交集靶点,应用Cytoscape v3.9.1软件构建蛋白质相互作用网络图,并筛选出核心靶点进行基因功能富集分析、信号通路分析和分子对接;通过基因鉴定将5月龄的早老素1/2条件性双基因敲除(PS cDKO)小鼠给予Pue(100 mg/kg)的腹腔注射治疗,然后进行行为学测试,测试结束后取材进行分子生物学检测。结果通过筛选获得Pue干预AD的核心靶点共118个。基因本体论(GO)功能富集分析得到148个条目(P<0.05),其中生物过程条目100个,细胞组成条目23个,分子功能条目25个;京都基因和基因组百科全书数据库(KEGG)通路分析富集得到11条信号通路(P<0.05),主要包括AD信号通路、磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/AKT)信号通路和丝裂原活化蛋白激酶(MAPK)信号通路等。分子对接结果显示,Pue与丝氨酸/苏氨酸蛋白激酶1(AKT1)、半胱氨酸蛋白酶3(CASP3)、白蛋白(ALB)、表皮生长因子受体(EGFR)、肿瘤坏死因子(TNF)、血管内皮生长因子A(VEGFA)可以对接。行为学结果显示,Pue可以改善PS cDKO小鼠受损的空间记忆(P<0.05)。免疫印迹(Western blot)结果显示,Pue下调了PS cDKO小鼠前额皮质中细胞外调节蛋白激酶(Erk1/2)的磷酸化水平的表达,并抑制了裂解的半胱氨酸蛋白酶-3(Cleaved Caspase-3)的表达水平。结论基于网络药理学和实验研究表明,Pue可以通过多靶点、多通路发挥对AD的干预作用,在临床和新药研发中具有参考价值。
Objective To investigate the molecular mechanism of action of Puerarin(Pue)in intervening Alzheimer's disease(AD)based on network pharmacology and molecular docking methods,and conduct animal experimental verification.Methods Use Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Integrated Pharmacophore Matching Platform(PharmaMapper),etc,to obtain the target of Pue.Use The Human Gene Database(GeneCards)to obtain AD related targets and intersection targets after mapping.Apply Cytoscape v3.9.1 to construct protein interaction network diagrams,and screen out core targets for GO enrichment analysis,KEGG signaling pathway analysis and molecular docking.The 5-month-old progerin-1/2 conditional double knockout(PS cDKO)mice were treated with intraperitoneal injection of Pue(100 mg/kg)based on gene identification,followed by behavioral testing,and after that the sample material was taken for molecular biological testing.Results A total of 118 core targets for Pue intervention in AD were obtained through screening.GO functional enrichment analysis revealed 148 items(P<0.05),including 100 items for biological process,23 items for cell composition,and 25 items for molecular function.KEGG pathway analysis enriched 11 signaling pathways(P<0.05),mainly including Alzheimer's disease signaling pathway,phosphatidylino-sitol 3-kinase/protein kinase B(PI3K/AKT)signaling pathway and mitogen-activated protein kinase(MAPK)signaling pathway,etc.Molecular docking results showed that Pue had better docking activity with AKT Serine/Threonine Kinase 1(AKT1),cysteine proteinase 3(CASP3),albumin(ALB),epidermal growth factor receptor(EGFR),tumor necrosis factor(TNF),and vascular endothelial growth factor A(VEGFA).Behavioral results showed that Pue could improve the impaired spatial memory in PS cDKO mice(P<0.05).Western blot results showed that Pue down-regulated the expression of Erk1/2 phosphorylation in the prefrontal cortex of PS cDKO mice,and inhibited the expression of Cleaved Caspase-3.Conclusion B
作者
巴宗韬
高外毛
热爱拉·阿合力江
徐颖
王健
BA Zongtao;GAO Waimao;AHELIJIANG Reaila;XU Ying;WANG Jian(School of Rehabilitation,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;School of Integrative Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)
出处
《上海中医药杂志》
CSCD
2023年第9期33-41,共9页
Shanghai Journal of Traditional Chinese Medicine
基金
国家自然科学基金项目(82174003)。
关键词
阿尔茨海默病
葛根素
网络药理学
分子对接
作用机制
中药研究
Alzheimer's disease
puerarin
network pharmacology
molecular docking
mechanism of action
traditional Chinese herbal medicine research
