基于网络药理学结合实验验证探讨桔梗皂苷D抗鼻咽癌的作用机制  

Exploring the mechanism of platycodin D in inhibiting nasopharyngeal carcinoma based on network pharmacology combined with experimental validation

作　　者：雷本蓝 杨江平 陈艳芳 黄精纶 张希平 马灿芳 梁钢[2] 焦爱军[2] LEI Benlan;YANG Jiangping;CHEN Yanfang;HUANG Jinglun;ZHANG Xiping;MA Canfang;LIANGGang;JIAO Aijun(The Third Affiliated Hospital of Guangxi Medical University,Nanning 530031,China;School of Pharmacy,Guangxi Medical University,Nanning 530021,China;Cancer Research Institute of the Second Affiliated Hospital of Zhejiang University School of Medicine,Hangzhou 310009,China;Guangxi Wantong Pharmaceutical Co.,Ltd,Nanning 530003,China)

机构地区：[1]广西医科大学第三附属医院,南宁530031 [2]广西医科大学药学院,南宁530021 [3]浙江大学医学院附属第二医院肿瘤研究所,杭州310009 [4]广西万通制药有限公司,南宁530003

出　　处：《广西医科大学学报》2025年第1期65-73,共9页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81960737)。

摘　　要：目的:通过网络药理学方法与细胞实验技术,旨在深入探讨桔梗皂苷D抗鼻咽癌方面的作用机制。方法:通过Pubchem、PharmMapper、GeneCards数据库获取桔梗皂苷D及鼻咽癌对应靶点并取交集后输入DAVID数据库中进行GO和KEGG富集分析,通过STRING数据库和Cytoscape软件获取交集靶点蛋白互作网络(PPI)、“桔梗皂苷D—靶点—鼻咽癌—通路”网络模型,筛选获得相关信号通路及核心靶点,对排名前5的核心靶点和桔梗皂苷D进行分子对接。利用CCK-8法、平板克隆实验、伤口愈合实验、Calcein-AM/PI染色分别评估桔梗皂苷D对鼻咽癌细胞增殖、克隆形成、迁移和凋亡的影响,western blotting检测MAPK/ERK通路相关蛋白表达水平。结果:网络药理学分析获得潜在治疗靶点115个,GO、KEGG富集分析获得生物过程309个、细胞组分44个、和分子功能83个,信号通路125个,综合PPI、“药物—靶点—鼻咽癌—通路”网络模型以及KEGG富集结果筛选获得MAPK1、HRAS、GRB2、MAPK8、MAPK10等10个核心靶点,其中MAPK1度值最高。分子对接结果显示,桔梗皂苷D与核心靶点有较好的结合活性。细胞实验结果显示,桔梗皂苷D可显著抑制鼻咽癌细胞的增殖、迁移及克隆形成能力,诱导凋亡,下调p-ERK蛋白表达(均P<0.05)。结论:桔梗皂苷D可能通过降低p-ERK蛋白的表达,进而抑制MAPK/ERK信号通路的激活,从而发挥抗鼻咽癌的作用。Objective:To delve into the underlying mechanism of platycodin D in inhibiting nasopharyngeal carcinoma(NPC)through a combination of network pharmacology and cellular experimental techniques.Methods:The corresponding targets of platycodin D and nasopharyngeal carcinoma were obtained from Pubchem,Pharm‐Mapper,and GeneCards databases,and their intersection was input into the DAVID database for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.The protein-protein interaction(PPI)network of the intersection targets and the“platycodin D-target-nasopharyngeal carcinoma-pathway”network model were obtained using the STRING database and Cytoscape software.Relevant signaling pathways and core targets were screened and obtained,and molecular docking was performed between the top five core targets and platycodin D.Cell counting kit-8(CCK-8)assay,plate cloning experiment,wound healing experiment,and Calcein-AM/PI staining were used to evaluate the effects of platycodin D on the proliferation,colony formation,migration and apoptosis of nasopharyngeal carcinoma cells.Western blotting was employed to verify the expression levels of proteins related to the MAPK/ERK pathway.Results:Network pharmacology analysis identified 115 potential therapeutic targets.GO and KEGG enrichment analyses yielded 309 biological processes,44 cellular components,83 molecular functions,and 125 signaling pathways.By integrating PPI,drug-target-diseasepathway network models and KEGG enrichment results,10 core targets including MAPK1,HRAS,GRB2,MAPK8,and MAPK10 were identified,with MAPK1 having the highest degree value.The molecular docking results showed that platycodon D had good binding activity with the core target.Cellular experiments revealed that platycodin D significantly inhibited NPC cell proliferation,migration,and colony formation,while inducing apoptosis and down-regulating the expression of p-ERK protein(P<0.05).Conclusion:Platycodin D may exert its anti-NPC effects by inhibiting the activation of the

关 键 词：桔梗皂苷D 鼻咽癌 网络药理学 MAPK/ERK信号通路 

分 类 号：R969[医药卫生—药理学]