中药新药与临床药理2025，Vol.36Issue(11)：1902-1913,12.DOI:10.19378/j.issn.1003-9783.2025.11.009
基于网络药理学及体外实验探讨蒲葵子治疗脑胶质瘤的作用机制
Exploring the Mechanism of Livistona chinensis Seeds in Treating Glioma Based on Network Pharmacology and In Vitro Experiments
摘要
Abstract
Objective To investigate the potential mechanism of Livistona chinensis seeds in the treatment of glioma using network pharmacology and in vitro experiments.Methods(1)Potential targets of Livistona chinensis seeds were obtained through literature review and the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).Glioma-related targets were collected from the GeneCards database,and the intersection was taken to identify potential therapeutic targets.Cytoscape 3.9.1 and the STRING database were used to construct an"herb-active ingredient-target"network and a protein-protein interaction(PPI)network,respectively,to screen key active ingredients and core targets.GO functional and KEGG pathway enrichment analyses of the intersection targets were performed using the R package,and molecular docking was conducted to validate interactions between key active ingredients and core targets.(2)The CCK-8 assay was used to determine the intervention concentrations of different polar fractions(n-butanol,ethyl acetate,petroleum ether)of the ethanol extract of Livistona chinensis seeds on U251 glioma cells.Based on this,U251 cells were divided into control,n-butanol(101.2 μg·mL-1),ethyl acetate(107.0 μg·mL-1),and petroleum ether(97.0 μg·mL-1)groups.Apoptosis was assessed using Annexin V-FITC flow cytometry;reactive oxygen species(ROS)levels were measured by fluorescence assay;RT-qPCR was used to detect mRNA expression of proto-oncogenes MYC,JUN,vascular endothelial growth factor A(VEGFA),FOS,epidermal growth factor(EGF),hypoxia-inducible factor 1α(HIF-1α),and Caspase-9;Western Blot was employed to measure protein expression of VEGFA,phosphorylated protein kinase B(p-AKT),HIF-1α,and Caspase-9.Results(1)A total of 17 active ingredients and 167 potential targets of Livistona chinensis seeds were identified.Key active ingredients included apigenin,emodin,β-sitosterol,isorhamnetin,and dehydrocavidine,while key targets included MYC,JUN,VEGFA,FOS,EGF,and HIF-1α.GO enrichment analysis revealed involvement in processes such as precursor metabolite and energy generation,mitochondrial protein complex formation,and guanosine nucleotide binding.KEGG analysis indicated that the treatment of glioma with Livistona chinensis seeds primarily involved pathways related to metabolism,cancer formation,and human T-cell leukemia virus 1 infection.Molecular docking results demonstrated strong binding affinity between key active ingredients and core targets.(2)The intervention concentrations for the n-butanol,ethyl acetate,and petroleum ether groups were determined to be 101.2,107.0,and 97.0 μg·mL-1,respectively.The proliferation indices of all treatment groups were significantly lower than those of the control group(P＜0.05,P＜0.01),while ROS relative fluorescence intensity was increased(P＜0.01).Apoptosis rates were significantly elevated in the n-butanol and ethyl acetate groups(P＜0.01),with an increasing trend in the petroleum ether group that was not statistically significant(P＞0.05).mRNA expression of JUN,VEGFA,and HIF-1α,as well as protein expression of VEGFA and HIF-1α,were significantly decreased in the n-butanol,ethyl acetate,and petroleum ether groups compared to the control(P＜0.05,P＜0.01),while mRNA and protein expression of Caspase-9 were increased(P＜0.01).mRNA expression of MYC and FOS,protein expression of p-AKT,and mRNA expression of EGF in the n-butanol group were significantly reduced(P＜0.05,P＜0.01).Although decreasing trends were observed in MYC,FOS,and EGF mRNA expression and p-AKT protein expression in the petroleum ether group,as well as EGF mRNA in the ethyl acetate group,these changes were not statistically significant(P＞0.05).Conclusion Livistona chinensis seeds exert anti-glioma effects through multiple components,targets,and pathways.The mechanism may be related to the modulation of key targets such as MYC,JUN,VEGFA,FOS,EGF,HIF-1α,and Caspase-9,as well as the regulation of critical signaling pathways including PI3K/AKT and HIF-1α/VEGF-mediated angiogenesis.关键词
脑胶质瘤/蒲葵子/网络药理学/分子对接/血管内皮生长因子A/PI3K/AKT/HIF-1α/VEGFKey words
glioma/Livistona chinensis seeds/network pharmacology/molecular docking/vascular endothelial growth factor A/PI3K/AKT/HIF-1α/VEGF分类
医药卫生引用本文复制引用
谢才军,李伟杰,罗文珍,侯丽,陈金曼,黄天华,张志强..基于网络药理学及体外实验探讨蒲葵子治疗脑胶质瘤的作用机制[J].中药新药与临床药理,2025,36(11):1902-1913,12.基金项目
广东省科学技术厅基金项目(粤科智字[2022]6号). （粤科智字[2022]6号）
