基于网络药理学的健脾解毒方抗结直肠癌的作用机制研究

  目的  基于网络药理学和分子对接研究健脾解毒方的抗结直肠癌作用机制。  方法  运用TCMSP、TCMID、ETCM数据库预测健脾解毒方有效成分及其作用靶点, 利用GeneCards数据库检索肿瘤相关靶点, 并进行药物-疾病靶点匹配。将共同靶点导入Cytoscape 3.7.2构建PPI网络并进行网络拓扑分析筛选关键靶点。通过R 3.6.3软件对共同靶点进行GO富集分析及KEGG通路富集分析。利用AutoDock平台进行分子对接, 预测有效成分与关键靶点的结合度, 并对关键靶点进行实验验证。  结果  共筛选出健脾解毒方有效成分24种, 肿瘤相关靶点86个, 其中关键靶点为PTGS2、HSP90AA1、PRSS1, 且与主要有效成分槲皮素、山柰酚均具有良好的结合活性。GO富集分析和KEGG通路富集分析提示健脾解毒方可能通过PI3K-Akt信号通路、MAPK信号通路发挥抗结直肠癌作用。实验验证发现健脾解毒方能够下调PTGS2、p38MAPK蛋白及mRNA表达, 而沉默PTGS2基因后健脾解毒方对下游基因p38MAPK的表达无明显调控作用, 且对结肠癌细胞转移能力无明显影响。  结论  健脾解毒方能够抑制结肠癌转移, 其机制与PTGS2介导的p38MAPK信号通路相关。 

Study on the Anti-Colorectal Cancer Mechanism of Jianpi Jiedu Decoction Based on Network Pharmacology

OBJECTIVE To explore the anti-tumor mechanism of Jianpi Jiedu Decoction(JPJDD)based on network pharmacology and molecular docking.METHODS TCMSP,TCMID and ETCM databases were used to predict the active ingredients and the targets of JPJDD,and GeneCards database was used to search tumor related targets and match the targets above.The common targets were imported into Cytoscape 3.7.2 to construct the PPI network and select the key targets by network topology analysis.GO enrichment analysis and KEGG pathway enrichment analysis were performed by R 3.6.3 software.Finally the Autodock 2.4 was used for molecular docking to predict the binding degree of the active ingredients to target genes.In addition,the key targets were verified by cell experiments.RESULTS 24 kinds of active ingredients and 86 tumor related targets were identified,including the key targets of PTGS2,HSP90AA1 and PRSS1,which were obtained by PPI network topology analysis.The molecular docking showed that all of the key targets had good binding activity with quercetin and kaempferol.GO enrichment analysis and KEGG pathway enrichment analysis suggested that JPJDD may play the role of anti-tumor through PI3K/Akt signal pathway and MAPK signal pathway.The results showed that JPJDD inhibited both the protein and mRNA expressions of PTGS2 and p38MAPK,but after silencing PTGS2 gene,the expression of downstream gene p38MAPK and the effect on colorectal cancer cells metastasis had no obvious response to JPJDD.CONCLUSION JPJDD can inhibit the metastasis of colorectal cancer via PTGS2 mediated p38MAPK signaling pathway.