基于系统药理学探究栀子苷改善阿尔茨海默病的作用机制及实验验证  

Mechanism and experiment verification of geniposide in improving Alzheimer’s disease based on system pharmacology

作　　者：崔珊珊 姜晶 薛慧 邱琦 刘宇欣 朴成玉[1] 雷霞 张亚峰[2] 张宁[1] CUI Shan-shan;JIANG Jing;XUE Hui;QIU Qi;LIU Yu-xin;PIAO Cheng-yu;LEI Xia;ZHANG Ya-feng;ZHANG Ning(School of Pharmacy,Heilongjiang University of Traditional Chinese Medicine,Harbin 150040;Wuxi Affiliated Hospital of Nanjing University of Traditional Chinese Medicine,Jiangsu Clinical Medical Innovation Center of Jiangsu Traditional Chinese Medicine Degenerative Osteoarthropathy,Wuxi Jiangsu 214071)

机构地区：[1]黑龙江中医药大学药学院,哈尔滨150040 [2]南京中医药大学无锡附属医院,江苏省中医退行性骨关节病临床医学创新中心,江苏无锡214071

出　　处：《中南药学》2025年第4期997-1003,共7页Central South Pharmacy

基　　金：国家自然科学基金面上项目(No.82174007);江苏省中医退行性骨关节病临床医学创新中心资助项目(No.20210405);黑龙江中医药大学科研基金项目(No.201819)。

摘　　要：目的基于GEO芯片、网络药理学和分子对接技术探索栀子苷改善阿尔茨海默病(AD)的潜在作用机制,并应用体外实验进行初步验证。方法通过公开数据库GEO挖掘栀子苷治疗AD的关键基因,借助PharmMapper检索药物靶点信息,利用GEO、OMIM、DrugBank、GeneCards和TTD筛选AD靶点。通过Cytoscape构建“活性成分-靶点”可视化网络图,通过DAVID数据库进行GO、KEGG和分子对接分析,预测其作用机制。通过栀子苷干预Aβ25-35诱导大鼠嗜铬瘤细胞(PC12细胞)所构建的AD细胞模型,利用MTT法检测栀子苷的有效浓度,ELISA法检测栀子苷对炎症因子肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)的影响,并通过Western blot验证核心靶点和通路。结果网络药理学结果显示,栀子苷与AD共有119个交集靶点,涉及信号传导、细胞迁移的正向调节等生物过程和蛋白激酶活性、酶结合和ATP结合等分析功能,富集在AD、MAPK信号通路、雌激素信号通路和PI3K-Akt等信号通路。JNK1等关键靶点参与调控MAPK信号通路。分子对接结果表明,栀子苷与JNK1具有良好的结合亲和力。体外实验表明,栀子苷能够提高PC12细胞增殖率,抑制炎症因子TNF-α、IL-1β和IL-6的释放,改善Aβ25-35对PC12细胞的损伤,提高ERβ的水平,降低p-JNK/JNK的水平。结论栀子苷可能通过增加ERβ的表达,抑制p-JNK/JNK的活性,进而发挥对Aβ损伤PC12细胞的保护作用。Objective To preliminarily verify the potential mechanism of geniposide in improving Alzheimer’s disease(AD)by in vitro experiments,based on chip mining,network pharmacology,and molecular docking.Methods Key genes modulated by geniposide in AD were initially mined from the Gene Expression Omnibus(GEO)database,followed by prediction of direct drug targets with PharmMapper.AD-related targets were concurrently curated from GEO,OMIM,DrugBank,GeneCards,and TTD databases,with overlapping candidates selected for subsequent analysis.A“compound-target”interaction network was constructed via Cytoscape to visualize relationships and prioritize hub targets based on topological parameters(degree centrality,betweenness centrality).Functional annotation via DAVID revealed significant enrichment in Gene Ontology(GO)biological processes and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways,while molecular docking simulations validated binding affinities between geniposide and core targets.Experimental validation involved treating Aβ25-35-induced PC12 cell models of AD with geniposide,followed by MTT assay to determine optimal concentrations,ELISA to quantify inflammatory cytokines(TNF-α,IL-1β,and IL-6),and Western blot analysis to confirm expression changes in key signaling pathways.Results Network pharmacology showed that geniposide mainly acted on key targets such as JNK1,and participated in the regulation of MAPK signaling pathway.Molecular docking showed that geniposide had good binding affinity to JNK1.In vitro experiments showed that geniposide increased the proliferation rate of PC12 cells,inhibited the release of inflammatory factors TNF-α,IL-1βand IL-6,improved the damage of Aβ25-35 to PC12 cells,increased the expression of ERβ,and decreased the expression of p-JNK/JNK.Conclusion Geniposide may inhibit the activity of p-JNK/JNK by increasing the expression of ERβ,thereby protecting PC12 cells damaged by Aβ.

关 键 词：栀子苷 阿尔茨海默病 PC12细胞 AΒ p-JNK1/JNK1 

分 类 号：R286[医药卫生—中药学]