基于网络药理学和体外实验研究黄连碱通过诱导铁死亡抑制鼻咽癌的机制  

Mechanism of Coptisine-Induced Ferroptosis in Inhibiting Nasopharyngeal Carcinoma:Evidence from Network Pharmacology and In Vitro Experiments

作　　者：王滨亚 陶阳阳 黄可盈 肖佳乐 何骁倩如 周赛男[1,2] 何迎春[1,2] 蔺婷 WANG Binya;TAO Yangyang;HUANG Keying;XIAO Jiale;HE Xiaoqianru;ZHOU Sainan;HE Yingchun;LIN Ting(Medcal School,Hunan University of Chinese Medicine,Changsha Hunan 410208,China;Hunan Key Laboratory for Prevention&Treatment of Ophthalmology and Otolaryngology Diseases with Chinese Medicine,Changsha Hunan 410208,China)

机构地区：[1]湖南中医药大学医学院,湖南长沙410208 [2]湖南中医药大学中医药防治眼耳鼻咽喉疾病湖南省重点实验室,湖南长沙410208

出　　处：《中医药导报》2025年第3期30-36,50,共8页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖南省自然科学基金资助项目(2023JJ30449);国家自然科学基金资助项目(82405496);湖南省自然科学基金-高校联合项目(2025JJ90022);湖南省大学生创新课题基金项目(S202310541114);湖南省大学生创新课题基金项目(S202310541100)。

摘　　要：目的:探讨黄连碱对鼻咽癌细胞5-8F与6-10B的铁死亡诱导效应及其作用机制。方法:运用网络药理学方法筛选黄连碱与鼻咽癌的共同靶点,构建蛋白质互作网络(PPI),通过基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析预测其潜在作用机制。将筛选所得靶点与铁死亡相关基因进行交集分析,并采用分子对接研究黄连碱与关键靶点的结合能力。随后采用MTT法评估黄连碱对鼻咽癌细胞的增殖抑制作用,并测定细胞内谷胱甘肽(GSH)、活性氧(ROS)和丙二醛(MDA)水平。采用Western blotting检测铁死亡关键蛋白谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族7成员11(SLC7A11)和信号转导与转录激活因子3(p-STAT3)的表达。结果:网络药理学分析显示,黄连碱与鼻咽癌共有150个共同靶点,其中热休克蛋白90α1(HSP90AA1)、热休克蛋白90β1(HSP90AB1)、信号转导与转录激活因子1(STAT1)、核因子κB1(NF-κB1)和前列腺素内酰胺合酶2(PTGS2)为关键靶点。KEGG富集分析发现趋化因子信号通路显著富集,提示STAT3信号通路可能参与调控铁死亡过程。PTGS2、核因子E2相关因子2(NFE2L2)等靶点为调控铁死亡关键靶点。分子对接结果显示黄连碱与PTGS2、NFE2L2具有良好的结合亲和力,结合能分别为-7.46、-6.76 kcal/mol(1 cal=4.2 J)。黄连碱能抑制鼻咽癌细胞的增殖活性,同时诱导鼻咽癌细胞内ROS和MDA水平升高,GSH水平下降,表明其具有诱导铁死亡的作用。Western blotting验证结果显示,黄连碱能降低鼻咽癌细胞GPX4、SLC7A11和p-STAT3蛋白表达水平,进一步证实了黄连碱通过调控STAT3信号通路诱导铁死亡的作用机制。结论:黄连碱能诱导鼻咽癌细胞5-8F与6-10B铁死亡,其可能机制为抑制STAT3信号通路。Objective:To investigate the ferroptosis-inducing effect of Coptisine(COP)on nasopharyngeal carcinoma(NPC)cells 5-8F and 6-10B,as well as the underlying mechanism.Methods:Network pharmacology approach was first performed to identify common targets between COP and NPC,and protein-protein interaction(PPI)network was constructed.Potential mechanisms were predicted through Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.The screened targets were intersected with ferroptosis-related genes,and molecular docking was performed to evaluate the binding affinity between COP and key targets.Subsequently,MTT assay was employed to evaluate the antiproliferative effects of COP on NPC cells.The ferroptosis-inducing effects were preliminarily investigated by measuring intracellular levels of glutathione(GSH),reactive oxygen species(ROS)and malondialdehyde(MDA)using commercial assay kits.Finally,Western blotting analysis was conducted to examine the protein expression levels of key ferroptosis regulators,including glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11)and phosphorylated signal transducer and activator of transcription 3(p-STAT3)to further elucidate the mechanism of COP-induced ferroptosis in NPC cells.Results:Network pharmacology analysis identified 150 common targets between COP and NPC,with heat shock protein 90 alpha family class a member 1(HSP90AA1),heat shock protein 90 alpha family class b member 1(HSP90AB1),signal transducer and activator of transcription 1(STAT1),nuclear factor kappa b subunit 1(NF-κB1),and prostaglandin-endoperoxide synthase 2(PTGS2)as key targets.KEGG enrichment analysis revealed significant enrichment in chemokine signaling pathway,suggesting STAT3 signaling pathway's involvement in ferroptosis regulation.Intersection analysis with ferroptosis-related genes identified PTGS2 and NFE2L2 as crucial regulators.Molecular docking revealed strong binding affinities between COP and PTGS2 and NFE2L2,with binding energies of-7.46 and-6.76 k

关 键 词：鼻咽癌 黄连碱 铁死亡 网络药理学 STAT3信号通路 

分 类 号：R285.5[医药卫生—中药学]