基于网络药理学结合细胞和分子实验探究地榆皂苷Ⅱ抗鼻咽癌的潜在靶点和作用机制

Potential targets and mechanism of Ziyuglycoside Ⅱ against nasopharyngeal carcinoma based on network pharmacology combined with cell and molecular experiments

目的:采用网络药理学的方法分析预测出地榆皂苷Ⅱ(ZiyuglycosideⅡ)治疗鼻咽癌(NPC)的相关作用靶点及通路,并通过实验进行验证。方法:通过Pubchem、Swiss Target Prediction、GeneCards等数据库查询获得地榆皂苷Ⅱ治疗鼻咽癌的相关作用靶点,并运用David数据库对靶点进行基因本体(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析,最后构建出"药物—靶点—疾病—通路"关系网络模型图。通过CCK-8细胞毒性实验探究地榆皂苷Ⅱ对鼻咽癌5-8F细胞增殖的影响。通过蛋白质印迹法(Western blotting)对预测出的靶点及其涉及到的信号通路进行验证。结果:网络药理学结果预测显示,地榆皂苷Ⅱ治疗鼻咽癌的潜在靶点有38个,GO功能注释和KEGG通路富集显示其机制可能与调控癌症信号通路(pathway in cancer)、磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B(AKT)信号通路、癌症蛋白聚糖通路(proteoglycans in cancer)、叉头转录因子O亚族(FOXO)信号通路等有关。实验结果显示,地榆皂苷Ⅱ对鼻咽癌5-8F细胞的增殖有明显的抑制作用,地榆皂苷Ⅱ可明显下调AKT蛋白和PI3K蛋白水平(P<0.05)。结论:地榆皂苷Ⅱ可有效抑制鼻咽癌5-8F细胞的增殖,其抗鼻咽癌机制可能与抑制PI3K/AKT信号通路活性有关。

Objective:To analyze and predict the target and pathway of Ziyuglucoside II in the treatment of nasopharyngeal carcinoma(NPC) by network pharmacology,and verified by experiments.Methods:The related targets of Ziyuglycoside II in the treatment of NPC were obtained by searching PubChem,Swiss target prediction,genecards and other databases,and the gene ontology(GO) function enrichment analysis and Kyoto Encyclopedia of genes and genomes(KEGG) pathway enrichment analysis were carried out by using David database,Finally,the network model of "drug targets disease pathway" is constructed.CCK-8 test was used to explore the inhibitory effect of Ziyuglycoside II on the proliferation of NPC 5-8F cells.Western blotting was used to detect the protein expressions of the predicted targets and related signaling pathways.Results:According to the results of network pharmacology,there are 38 potential targets of Ziyuglycoside II in the treatment of NPC.GO functional annotation and KEGG pathway showed it might be related to the regulation of the signaling pathway in cancer,phosphatidylinositol-3-kinases(PI3 K)/protein kinase B(Akt) signaling pathway,proteoglycans in cancer pathway,forkhead transcription factor of the O class(FOXO) signaling pathway and so on.Ziyuglycoside II significantly inhibited the proliferation of NPC 5-8 F cells and down-regulated the protein expressions of Akt and PI3 K(P<0.05).Conclusion:Ziyuglycoside II can effectively inhibit the proliferation of NPC 5-8 F cells and its anti NPC mechanism may be related to the inhibition of PI3 K/Akt signal pathway activity.