基于网络药理学探索梅花草提取物抑制脑胶质瘤的作用机制

Exploring the mechanism of Meihuacao extracts in inhibiting glioma based on network pharmacology and preclinical experiments

目的运用网络药理学技术结合细胞及动物实验探讨梅花草提取物抑制脑胶质瘤恶性增殖的作用机制。方法通过中药系统药理学分析平台(TCMSP)数据库筛选梅花草的有效化合物,使用OMIM、Genecards、Pubchem等数据库预测梅花草有效化合物对脑胶质瘤的潜在作用靶点,应用Cytoscape 3.9.1和STRING 11.5数据库构建化合物-靶点网络、蛋白质互作(PPI),筛选出核心基因,运用DAVID 6.8数据库对靶点进行基因本体(GO)的富集分析和京都基因与基因组百科全书(KEGG)的通路富集分析,并在细胞和动物水平评价相关信号通路和表型变化。结果梅花草含有14个靶点活性成分,包括Celahin B、Isoboonein、槲皮素(Quercetin)、山奈酚(Kaempferol)、没食子酸、Mussaenin A、金丝桃苷、山奈酚-3-O-β-芸香糖苷(Nicotiflorin)、山奈酚3-O-葡萄糖苷(Astragalin)、槲皮素3-O-葡萄糖苷(Isoquercitrin)、槲皮素7-O-葡萄糖苷(Quercimeritrin)、山奈酚-7-O-葡萄糖苷(Kaempferol 7-O-glucoside)、槲皮素-3,7-二-O-葡萄糖苷(Quercetin 3,7-diglucoside)、皂苷(Saponin),化合物对应的靶点338个;通过韦恩图将梅花草有效化合物与肿瘤相关的334个关键靶点基因利用DAVID在线分析工具进行肿瘤相关靶点的GO与KEGG分析。GO分析结果表明,梅花草提取物抗脑肿瘤的作用机制可能与炎症状态、缺氧反应、细胞增殖与凋亡等有关;KEGG分析结果发现梅花草有效化合物作用的靶点基因与PI3K/AKT、MAPK、TNF、HIF-1α等抗肿瘤信号通路息息相关,并在细胞及动物水平获得肿瘤的抑制表型和线粒体相关PI3K/AKT关键信号通路的验证。结论梅花草提取物显著抑制脑胶质瘤细胞的恶性生长,呈现浓度依赖性,其发挥作用可能与PI3K/AKT、线粒体代谢等信号通路有关。

Objective To explore the mechanism of Meihuacao extracts in inhibiting the malignant proliferation of gliomas by using network pharmacology techniques combined with cell and animal experiments.Methods We screened the effective compounds of Meihuacao extracts through the Traditional Chinese Medicine System Pharmacology Analysis Platform(TCMSP)database,predicted the potential targets of effective compounds of Meihuacao on gliomas using OMIM,Genecards,Pubchem and other databases,and constructed the compound-target network by applying Cytoscape 3.9.1 and STRING 11.5 databases,Protein Interaction(PPI),screened the core genes,and used DAVID 6.8 database to perform Gene Ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis on the targets,and evaluated the related signaling pathways and phenotypes at cellular and animal levels.Results Meihuacao extracts contain 14 target active ingredients,including Celahin B,Isoboonein,Quercetin,Kaempferol,Gallic acid,Mussaenin A,Hypericin,Kaempferol-3-O-β-rutinoside(Nicotiflorin),Kaempferol 3-O-glucoside(Astragalin),Quercetin 3-O-glucoside(Isoquercitrin),Quercetin 7-O-glucoside(Quercimeritrin),Kaempferol 7-O-glucoside,Quercetin 3,7-di-O-glucoside(Quercetin 3,7-diglucoside),Saponin,compounds corresponding to 338 related targets;The 334 key target genes of effective compounds related to cancer in Meihuacao extracts using Venn analysis were subjected to GO and KEGG analysis of cancer-related targets using DAVID online analysis tool.The GO analysis showed that the anti-glioma of Meihuacao extracts.The mechanism may be related to the inflammatory status,hypoxia response,cell proliferation and apoptosis,et al.KEGG analysis revealed that the active substances of Meihuacao extracts were closely related to the anti-tumor signaling pathways,such as PI3K/AKT,MAPK,TNF,HIF-1α,et al.,and phenotypic inhibitory effects and validation of the key signaling pathways of mitochondrial related PI3K/AKT were obtained at the cellular and animal levels.Conclusion Meihuacao extracts significantly inhibited the malignant growth of glioma cells,showing concentration-dependence,and its action may be related to PI3K/AKT,mitochondrial metabolism and other signaling pathways.