莪术抗肿瘤活性成分的网络药理学研究

Study on anti-tumor active ingredients of Curcuma zedoaria by network pharmacological

目的基于网络药理学,探究中药莪术的主要活性成分及其抗肿瘤的作用机制。方法在中药系统药理学分析平台(TCMSP)数据库中检索莪术的化学成分,根据“类药五原则”和“口服生物利用度”≥30%的标准筛选莪术的活性成分,并预测其活性成分所对应的作用靶点,采用Cytoscape 3.7.2软件构建莪术活性成分-预测靶点网络图。在Genecards数据库中以“tumor”为关键词搜索抗肿瘤的靶点,与TCMSP数据库的莪术靶点映射,筛选出共同靶点作为莪术抗肿瘤的作用靶点。将莪术抗肿瘤靶点导入STRING数据库中进行蛋白质-蛋白质相互作用分析,构建靶蛋白之间相互作用的网络图(PPI)。利用Cytoscape 3.7.2的“CytoNCA”插件筛选莪术的抗肿瘤的核心靶点,通过RCSB数据库找到对应蛋白亚型的PDB-ID,用于下一步的分子对接。利用Discovery studio 3.1软件进行分子对接。最后,借助Cytoscape 3.7.2软件绘制莪术潜在活性成分与抗肿瘤作用靶点的网络图。使用Metascape数据库对莪术抗肿瘤靶点进行KEGG信号通路和GO生物过程的富集分析。结果获得莪术中口服吸收良好的类药活性成分43种,对应靶点72个。由富集分析结果可知:莪术抗肿瘤作用主要涉及鞘磷脂信号通路、凋亡信号通路的负调控和神经酰胺通路,以及低氧反应、活性氧代谢、T细胞激活、核外雌激素信号、血清素激活、蛋白质复合物齐聚等过程,并得到了7个核心靶点。分子对接结果显示:最佳候选分子双去氧基姜黄素(cop.43)可以和蛋白3ENE稳定地结合并与氨基酸残基LYS807、LYS883、GLU880等发生氢键作用,与TYR867发生Pi-Pi共轭。结论采用网络药理学和分子对接联用的方法筛选莪术治疗肿瘤的潜在活性成分和抗肿瘤的作用靶点,揭示了莪术治疗肿瘤具有多靶点、多通路的潜在作用机制,为莪术抗肿瘤的药效物质基础和作用机制的研究奠定了基础。

OBJECTIVE To explore the main active ingredients and anti-tumor mechanism of Curcuma zedoaria based on network pharmacology.METHODS Chemical constituents of C.zedoaria were obtained through TCMSP database,and the active ingredients of C.zedoaria were screened according to oral bioavailability(≥30%)and Lipinski's rule of five,and their corresponding targets were predicted.Cytoscape 3.7.2 software was used to construct the active ingredients-predicted targets network of C.zedoaria.In Genecards database,"tumor"was used as the key word to search for anti-tumor targets,and the common targets were selected by mapping with the predicted targets of C.zedoaria,which will be regarded as the anti-tumor targets of C.zedoaria.The anti-tumor targets of C.zedoaria were imported into the STRING database for protein-protein interaction analysis,and the protein-protein interaction network was built.The core anti-tumor targets of C.zedoaria were screened by plug-in CytoNCA in Cytoscape 3.7.2 software,and the PDB-IDs of the corresponding proteins were found by searching for the RCSB database,which could be used for the molecular docking in the next stage.Discovery Studio 3.1 software was used to perform molecular docking.Finally,Cytoscape 3.7.2 software was used to construct the active ingredients-anti-tumor targets network of C.zedoaria.GO and KEGG enrichment analyses of anti-tumor targets of C.zedoaria were performed using the Metascape database.RESULTS Forty-three active ingredients with good oral absorption and drug-like properties were screened from C.zedoaria,with seventy-two corresponding targets.The results of the enrichment analyses showed that the anti-tumor effects of C.zedoaria mainly involved sphingolipid signaling pathway,negative regulation of apoptotic signaling pathway,ceramide pathway,hypoxic reaction,reactive oxygen metabolism,T cell activation,extra-nuclear estrogen signaling,serotonin activation,and protein complex oligomerization.Finally,7 core targets were obtained.The molecular docking results of the active ingredients from C.zedoaria showed that the best candidate molecule,bisdemethoxycurcumin(cop.43),could bind stably to protein 3ENE,and form H-bonds with residues LYS807,LYS883 and GLU880,and form pi-pi stacking interaction with TYR867.CONCLUSION In this study,the potential anti-tumor active ingredients from C.zedoaria and the corresponding anti-tumor targets are identified by using network pharmacology and molecular docking analysis,and it reveals that C.zedoaria has the characteristics of multi-target and multi-pathway in treating tumors,which lays a theoretical foundation for the further research on the pharmacodynamic material basis and mechanisms of C.zedoaria in treating tumor.