摘要
目的:运用网络药理学技术,并结合动物实验,探讨白头翁皂苷B4调节小鼠乳腺癌肿瘤免疫微环境(TIME)及抗肿瘤的作用机制。方法:采用网络药理学,利用相关数据库筛选白头翁调节TIME的活性成分,检索TIME相关靶点,将成分与靶点取交集,运用STRING数据库构建蛋白交互作用(PPI)网络,DAVID数据库进行基因本体(GO)和京都基因与基因组百科全书(KEGG)通路富集分析,Cytoscape软件构建成分-靶点-通路网络分析关键通路,以获得白头翁皂苷B4的预测作用靶标。建立乳腺癌动物模型,白头翁皂苷B4水溶液进行干预,通过流式细胞术检测肿瘤免疫相关细胞的含量,并采用Western Blot法检测肿瘤中STAT3蛋白的表达。结果:网络药理学分析显示,白头翁调节TIME的核心成分中白头翁皂苷B4主要作用于STAT3靶标基因。GO富集分析发现白头翁调节TIME参与的生物学过程主要有肽酪氨酸磷酸化、肽酪氨酸修饰等;KEGG富集通路结果显示,EGFR酪氨酸激酶抑制剂耐药性、内分泌抗药性、癌症中的蛋白多糖等富集程度较高。动物实验结果表明,白头翁皂苷B4水溶液在高、低2个剂量下均可显著抑制肿瘤生长,其中白头翁皂苷B4高剂量组减少Tregs细胞、TLR4^(+)细胞的含量作用更为明显。Western Blot实验发现,与模型组比较,经白头翁皂苷B4治疗瘤内STAT3蛋白表达显著下调。结论:白头翁皂苷B4可能通过抑制免疫抑制细胞的活性或数量,减少肿瘤的免疫逃逸和免疫抑制微环境,增强机体抗肿瘤免疫反应,主要作用于STAT3靶标基因,下调其瘤内蛋白表达,为其在肿瘤免疫治疗中的应用提供理论依据。
Objective:To explore the mechanism of anemoside B4 in regulating the tumor immune microenvironment(TIME)and its anti-tumor effect through network pharmacology techniques combined with animal experiments.Methods:With the help of network pharmacology,relevant databases were used to screen the active components of Pulsatilla chinensis that regulate TIME,and the TIMErelated targets were retrieved.The components and targets were intersected.The STRING database was used to construct a protein-protein interaction(PPI)network,the DAVID database was used for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis,and the Cytoscape software was used to construct a component-target-pathway network to analyze the key pathways,to obtain the predicted action targets of anemoside B4.A breast cancer animal model was established and intervened with an aqueous solution of anemoside B4.Flow cytometry was used to detect the content of tumor immunity-related cells,and the Western Blot method was used to detect the expression of STAT3 protein in tumors.Results:Network pharmacology analysis showed that among the core components of Pulsatilla chinensis that regulate TIME,anemoside B4 mainly acts on the STAT3 target gene.GO enrichment analysis found that the biological processes involved in the regulation of TIME by Pulsatilla chinensis mainly included peptide tyrosine phosphorylation,peptide tyrosine modification,etc.;the results of KEGG pathway enrichment showed that the enrichment degrees of EGFR tyrosine kinase inhibitor resistance,endocrine resistance,proteoglycans in cancer,etc.,were relatively high.The results of animal experiments showed that the aqueous solution of B4 at both high and low doses could significantly inhibit tumor growth,and the contents of regulatory T cells(Tregs)and TLR4^(+)cells in the tumor were significantly reduced for the high-dose anemoside B4 group.Western Blot analysis revealed that,compared with the model group,the intratumoral STAT3 protein expression was significantly downregulated in the anemoside B4 treated group.Conclusion:Anemoside B4 may inhibit the activity or number of immunosuppressive cells,reduce tumor immune escape and the immunosuppressive microenvironment,enhance the body's anti-tumor immune response,and mainly act on the STAT3 target gene,downregulating the STAT3 protein expression,providing a theoretical basis for its application in tumor immunotherapy.
作者
李翔
胡雪婉
查敏
冯育林
杨世林
张婧
LI Xiang;HU Xuewan;ZHA Min;FENG Yulin;YANG Shilin;ZHANG Jing(National Pharmaceutical Engineering Center for Solid Preparation in Chinese Herbal Medicine,Nanchang 330006,China;Key Laboratory of Modern Preparation of TCM,Ministry of Education,Nanchang 330004,China;State Key Laboratory for the Modernization of Classical and Famous Prescriptions of Chinese Medicine,Nanchang 330006,China)
出处
《江西中医药大学学报》
2026年第1期93-100,共8页
Journal of Jiangxi University of Chinese Medicine
基金
江西省青年井冈学者项目(QNJG2018077)。
