基于网络药理学吴茱萸治疗结直肠癌作用机制探讨及实验验证

Mechanism of Evodia rutaecarpa in treatment of colorectal cancer based on network pharmacology and experimental verification

目的 采用网络药理学、分子对接及实验验证的方法,探讨吴茱萸治疗结直肠癌的作用机制。方法 运用中药系统药理学数据库与分析平台(TCMSP)数据库及查阅国内外相关文献收集吴茱萸活性成分,通过GeneCards、OMIM、TTD数据库收集结直肠癌靶点,将成分-疾病靶点取交集后进行基因本体(GO)注释及京都基因与基因组百科全书(KEGG)通路富集分析。AutoDock vina对核心成分与核心靶点进行分子对接。通过细胞增殖与活性检测-8(CCK-8)法和流式细胞仪检测异鼠李素对SW480细胞增殖和凋亡的影响;通过蛋白免疫印迹法(Western blotting法)检测细胞中肿瘤蛋白p53(TP53)、丝氨酸/苏氨酸蛋白激酶1(AKT_(1))、血管内皮生长因子A(VEGFA)的蛋白表达情况。结果 吴茱萸30个活性成分对应174个靶点,结直肠癌对应1 484个靶点,二者取交集获得98个靶点,GO涉及细胞增殖、细胞凋亡等生物过程,KEGG富集为癌症通路、PI3K/AKT等信号通路,分子对接结果显示异鼠李素与TP53、AKT_(1)和VEGFA亲和力良好。与对照组比较,异鼠李素可抑制SW480细胞增殖,促进细胞凋亡;与对照组比较,异鼠李素下调了AKT_(1)和VEGFA的蛋白表达水平(P<0.01),上调TP53的蛋白表达水平(P<0.01)。结论 吴茱萸治疗结直肠癌可通过多成分、多靶点、多通路途径来发挥作用,其中异鼠李素是主要活性成分。

Objective To investigate the mechanism of Evodia rutaecarpa in treatment of colorectal cancer based on network pharmacology,molecular docking and experimental verification.Methods The active ingredients of E.rutaecarpa were collected by TCMSP and relevant domestic and foreign literature,targets to colorectal cancer were collected through GeneCards,OMIM and TTD databases.The component-disease intersection targets were analyzed through GO and KEGG analysis.AutoDock vina was used to molecular docking between the core components and the core targets.The effects of isorhamnetin on proliferation and apoptosis of SW480 cells by cell counting kit-8(CCK-8)and flow cytometry.Western blotting was employed to determine the expression levels of TP53,AKT_(1) and VEGFA.Results The 30 active components of E.rutaecarpa corresponding to 174 targets,1484 corresponding targets for colorectal cancer,and 98 targets for the intersection of the two.GO involve biological processes such as cell proliferation,apoptosis and so on.KEGG enrichment pathways involve cancer pathway,PI3K-Akt signaling pathway.Molecular docking results showed that isorhamnetin had a good affinity with TP53,AKT_(1),VEGFA.Isorhamnetin can inhibit proliferation of SW480 cell and and promote apoptosis compared with the group.Compared with the control group,isorhamnetin down-regulated the expression of AKT_(1) and VEGFA(P<0.01)and up-regulated the expression of TP53(P<0.01).Conclusion E.rutaecarpa can play a role in treatment of colorectal cancer through multi-component,multi-target and multi-pathway pathways,and isorhamnetin is the main active ingredient.