网络药理学方法探讨苦丁茶防治食管鳞癌的活性成分及分子机制的研究

目的:应用网络药理学技术探讨苦丁茶的主要活性成分并预测其防治食管鳞癌的分子作用机制。方法:通过检索中药系统药理学数据库和分析平台 (Traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP) 中苦丁茶所有成分数据，筛选出苦丁茶主要入血活性成分，应用Swiss target prediction预测主要成分的蛋白作用靶点；在GEO数据库中查找食管鳞癌相关基因；通过Cytoscape进行网络拓扑分析，构建苦丁茶主要活性成分的作用靶点与食管鳞癌相关基因的相互关系网络，筛选出苦丁茶防治食管鳞癌的关键靶点；采用STRING中GO功能分析和DAVIDE的KEGG信号通路富集，分析苦丁茶防治食管鳞癌的分子作用机制。结果:利用TCMSP 模型筛选出94 个苦丁茶有效成分中的6个主要活性成分：蒲公英赛醇（Taraxerol）、丁子香萜（Mairin)、β-谷甾醇（beta-sitosterol）、棕榈酸异戊酯（amyrin Palmitate）、儿茶素没食子酸酯（(-)-Catechin gallate）、坡模酸（tetradecahydropicene-4a-carboxylic acid）。应用Swiss target prediction软件预测出6个主要活性成分的174个蛋白作用靶点，活性成分与作用靶点之间形成392条对应关系。应用GEO数据库中检索食管鳞癌的差异表达基因，根据差异显著性，选取前250个差异表达基因；经网络拓扑分析后，筛选出73个苦丁茶活性成分靶点与食管鳞癌相关基因的关键交叉节点。应用STRING中GO功能富集和DAVIDE的KEGG通路富集分析结果显示，苦丁茶作用于食管鳞癌的关键节点涉及的生物过程主要包括：超分子纤维组织，细胞骨架组织，多细胞组织过程的调控等；分子功能主要包括：细胞骨架蛋白结合、蛋白质结合、酶结合、肌动蛋白结合、烟酰胺腺嘌呤二核苷酸(NAD)结合等；信号通路主要被富集于癌症中的蛋白多糖、环鸟苷酸蛋白激酶G信号通路、过氧化物酶体增殖物激活受体信号通路、脂肪细胞因子信号通路等。结论:通过网络药理学分析发现，苦丁茶的6个主要活性成分通过多个关键靶点基因、多个生物过程、多个分子功能及多条信号通路发挥防治食管鳞癌的作用。本研究分析结果表明苦丁茶的活性成分具有防治食管鳞癌的潜在功效。

Study on active components and molecular mechanisms of Ilex Kudingcha in the prevention and treatment of esophageal squamous cell carcinoma by network pharmacology

AIM: To investigate the main active components of Ilex Kudingcha and predict its molecular mechanism of action in the prevention and treatment of esophageal squamous cell carcinoma by network pharmacology. METHODS: The main active ingredients of Ilex Kudingcha were screened out by searching the data of all components of it in TCMSP. The protein targets of these main active ingredients were predicted by using Swiss target prediction. The Esophageal Squamous Cell Carcinoma (ESCC) related genes were analyzed by GEO Database. The topology analysis network of the interaction between the main active components of Ilex Kudingcha and the ESCC related genes was constructed by Cytoscape. Then the key targets of Ilex Kudingcha were dig out in the prevention and treatment of ESCC. The molecular mechanism of Ilex Kudingcha in the prevention and treatment of ESCC was analyzed by the the GO in STRING and the enrichment of KEGG signal pathway in DAVIDE. RESULTS:Six main active ingredients of 94 compounds of Ilex Kudingcha were screened by TCMSP model. There are Taraxerol, Mairin, beta-sitosterol, amyrin Palmitate, (-)-Catechin gallate and tetradecahydropicene-4a-carboxylic acid. The 174 protein targets of 6 main active ingredients were predicted. 392 corresponding relationships were formed between active ingredients and targets by Swiss target prediction. According to the difference significance, the top 250 differentially expressed genes of ESCC were selected in GEO database.73 key targets between active ingredients of Ilex Kudingcha and genes related to ESCC were dig out by topology analysis. The results of enrichment analysis of GO in STRING showed that the biological processes of Ilex Kudingcha on the key nodes of ESCC mainly included: Supramolecular fiber organization, cytoskeleton organization, regulation of multicellular organismal process, etc; the molecular functions mainly included: cytoskeleton protein binding, protein binding, enzyme binding, actin binding and NAD binding，etc. The results of enrichment analysis of KEGG pathway in DAVIDE showed that the signal pathways were mainly enriched in proteoglycan in cancer, cyclic guanosine protein kinase G signal pathway, oxidosome proliferator-activated receptor signaling pathway, adipocytokine signaling pathway, etc. CONCLUSION:Six main active ingredients of Ilex Kudingcha play a role in the prevention and treatment of ESCC through multiple key target genes, multiple biological processes, multiple molecular functions and multiple signaling pathways. The results indicated that the active ingredients of Ilex Kudingcha had potential efficacy in preventing and treating ESCC.