摘要
目的:基于GEO数据联合网络药理学筛选槲皮素改善阿尔茨海默病(AD)潜在靶点及通路,结合动物实验验证。方法:从GEO数据库筛选AD数据集,通过SwissTargetPrediction、GeneCards和DrugBank数据库筛选槲皮素作用靶点。构建交集靶点韦恩图,STRING数据库构建蛋白质-蛋白质相互作用(PPI)网络;Cytoscape软件筛选核心靶点,利用AutoDock Vina进行分子对接;DAVID数据库进行基因本体(GO)、京都基因和基因组百科全书(KEGG)通路富集分析。动物实验选用6周龄APP/PS1双转基因及正常C57BL/6雄性小鼠,分为野生型动物组(WT)、模型组(APP/PS1)和模型+槲皮素干预组(APP/PS1+Que)。Y迷宫检测小鼠学习记忆能力,Real time RT-PCR检测核心靶点mRNA表达水平,ELISA检测小鼠前额叶皮质中肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)和白介素-6(IL-6)的含量,Western blot检测磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)和核因子κB p65(NF-κB p65)通路蛋白磷酸化水平。结果:共获取二者交集靶标31个,其中CDK1、EGFR、HSP90AB1、PARP1和STAT3为核心靶点,分子对接提示槲皮素与核心靶点具有结合力,富集分析得到GO富集条目157条,KEGG通路86条;主要涉及AD途径、NF-κB和PI3K/AKT等通路。APP/PS1小鼠的行为学和分子生物学检测表明,槲皮素显著提高APP/PS1小鼠学习记忆能力,对核心靶点具有调控作用,能够降低前额叶皮质TNF-α、IL-1β和IL-6含量,促进PI3K/AKT通路蛋白磷酸化及抑制NF-κB p65通路蛋白磷酸化。结论:槲皮素可能通过PI3K/AKT和NF-κB p65通路抑制炎症因子的表达发挥抗AD作用,具有多靶点、多通路的特点。
Objective:To identify potential targets and pathways through which Quercetin may alleviate Alzheimer disease(AD)by integrating the Gene Expression Omnibus(GEO)databases and network pharmacology,which was veified through animal experiments.Methods:Alzheimer disease(AD)-related datasets were retrieved from the GEO database.Potential targets of Quercetin were predicted using the SwissTargetPrediction,GeneCards,and DrugBank databases.Venn diagram was constructed to identify overlapping targets between Quercetin and AD.Protein-protein interaction(PPI)network was established via the STRING database,and core targets were identified using Cytoscape software.Molecular docking analysis was performed using AutoDock Vina.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were conducted using the DAVID database.For in vivo experiments,six-week-old male APP/PS1 double transgenic mice and age-matched wild-type C57BL/6 mice were used.Animals were divided into three groups:WT group,APP/PS1 group,and APP/PS1+Quercetin(Que)group.The Y-maze test was employed to evaluate learning and memory abilities.The mRNA expression levels of core targets were measured by real time RT-PCR.ELISA was used to detect the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in the prefrontal cortex.Western blot was conducted to assess the phosphorylation levels of proteins in the phosphoinositide 3-kinase/protein kinase B(PI3K/AKT)and nuclear factor-κB p65(NF-κB p65)signaling pathways.Results:A total of 31 overlapping targets between quercetin and AD were identified,among which CDK1,EGFR,HSP90AB1,PARP1,and STAT3 were recognized as core targets.Molecular docking results indicated strong binding affinities between Quercetin and these core targets.Enrichment analysis yielded 157 GO terms and 86 KEGG pathways,primarily involving Alzheimer disease,NF-κB and PI3K/AKT signaling pathways.Behavioral and molecular biological assessments in APP/PS1 mice demonstrated that Quercetin significantly improved learning and memory abilities,exerted regulatory effects on core targets,reduced the levels of TNF-α,IL-1βand IL-6 in the prefrontal cortex,promoted phosphorylation of PI3K/AKT pathway proteins,and inhibited phosphorylation of NF-κB p65 pathway proteins.Conclusion:Quercetin may exert anti-AD effects by suppressing the expression of inflammatory cytokines through the PI3K/AKT and NF-κB p65 pathways.Its mechanism of action is characterized by multi-target and multi-pathway regulation.
作者
赵岩
王宗宝
柴雪洁
安朋朋
段贞
冯媛
李永征
ZHAO Yan;WANG Zongbao;CHAI Xuejie;AN Pengpeng;DUAN Zhen;FENG Yuan;LI Yongzheng(Qingdao Hiser Hospital Affiliated of Qingdao University/Qingdao Traditional Chinese Medicine Hospital,Qingdao 266000,China)
出处
《神经解剖学杂志》
北大核心
2025年第6期749-758,786,共11页
Chinese Journal of Neuroanatomy
基金
山东省中医药科技项目(M-2022010)。
关键词
槲皮素
阿尔茨海默病
生物信息
网络药理学
分子对接
小鼠
quercetin
Alzheimer disease(AD)
bioinformatics
network pharmacology
molecular docking
mouse
