天然产物研究与开发 ›› 2022, Vol. 34 ›› Issue (10): 1784-1793.doi: 10.16333/j.1001-6880.2022.10.017

• 数据研究 • 上一篇    下一篇

基于网络药理学、分子对接及实验验证探讨蚕梅方治疗结直肠癌的作用机制

周亚秋1,周红光1,2*   

  1. 1南京中医药大学第一临床医学院肿瘤研究所,南京 210046;2南京中医药大学附属医院,南京 210009
  • 出版日期:2022-10-28 发布日期:2022-10-31
  • 基金资助:
    国家自然科学基金面上项目(81973737);吴勉华全国名老中医药专家传承工作室(2022-75)

Mechanism of Canmei Decoction in the treatment of colorectal cancer based on network pharmacology and molecular docking

ZHOU Ya-qiu1,ZHOU Hong-guang1,2*   

  1. 1Cancer Institute,the First Clinical College of Nanjing University of Chinese Medicine,Nanjing 210046,China;2Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210009,China
  • Online:2022-10-28 Published:2022-10-31

摘要: 基于网络药理学、分子对接及实验验证探讨蚕梅方(Canmei Decoction,CMF)治疗结直肠癌潜在的作用机制。通过TCMSP数据库、化学成分数据库、文献挖掘等方法收集CMF的活性成分,经ADME筛选获得候选靶点,利用UniProt数据库查询靶点蛋白对应的人类基因;通过GeneCards、OMIM数据库收集有关结直肠癌的靶点,抽取交集网络获得候选基因。利用STRING数据库构建交集靶点蛋白相互作用网络(PPI),选出关键靶点基因。将共同靶基因进行GO功能富集与KEGG通路分析。使用分子对接技术对化合物核心成分与关键靶点进行对接验证,并对核心化学成分进行实验验证。体外实验验证:采用MTT、Annexin V-FITC/PI双染法和Western blot法行体外实验验证不同浓度槲皮素对HCT116细胞的抑制率和凋亡相关蛋白表达水平的影响。实验得到CMF所含化合物对应靶点166个、结直肠癌对应靶点1 028个,两者交集的关键靶点79个。关键靶点GO富集条目中生物过程相关的条目1 609条,细胞组成相关的条目41条,分子功能相关的条目123条;关键靶点的KEGG富集通路涉及癌症通路、PI3K-Akt信号通路、IL-17信号通路、细胞转录、细胞凋亡等。分子对接结果显示CMF核心成分与结直肠癌关键靶点亲和力良好。细胞实验结果显示,不同浓度的槲皮素干预HCT116细胞后可显著抑制细胞增殖和促进细胞凋亡,降低抗凋亡蛋白Bcl-2表达、增加促凋亡蛋白BAX表达。综上结果表明,CMF防治结直肠癌的潜在作用机制可能与调控免疫炎症反应、细胞自噬、细胞增殖及凋亡等过程有关,具有多靶点、多途径、多系统的机制特点。

关键词: 蚕梅方, 结直肠癌, 网络药理学, 作用机制, 槲皮素

Abstract:

To explore the potential mechanism of Canmei Decoction (CMF)in the treatment of colorectal cancer based on network pharmacology,molecular docking and experimental verification.The active components of CMF were collected by TCMSP database,chemical composition database,literature mining and other methods,and candidate targets were obtained through ADME screening,and the human genes corresponding to the target proteins were queried by UniProt database;targets related to colorectal cancer were collected through GeneCards and OMIM databases,and candidate genes were obtained by extracting the intersection network.Using STRING database,the protein-protein interaction network (PPI) of intersection target was constructed,and the key target genes were selected.The common target genes were enriched by GO function and analyzed by KEGG pathway.Molecular docking technology was used to verify the docking between the core components of the compound and the key targets,and the core chemical components were experimentally verified.In vitro validation:MTT,Annexin V-FITC/PI double staining and Western blot were used to verify the inhibitory rate of different concentrations of quercetin on HCT116 cells and the expression levels of apoptosis-related proteins.Experiments obtained 166 corresponding targets for compounds contained in CMF,1 028 corresponding targets for colorectal cancer,and 79 key targets for the intersection of the two.There were 1 609 items related to biological process,41 items related to cell composition and 123 items related to molecular function in the key target GO enrichment items.KEGG enrichment pathways of key targets involve cancer pathway,PI3K-Akt signaling pathway,IL-17 signaling pathway,cell transcription,apoptosis and so on.Molecular docking results showed that the core components of CMF had a good affinity with the key targets of colorectal cancer.The results of cell experiments showed that quercetin with different concentrations could significantly inhibit cell proliferation and promote cell apoptosis,reduce the expression of anti-apoptotic protein Bcl-2 and increase the expression of pro-apoptotic protein BAX.In conclusion,the potential mechanism of CMF in the prevention and treatment of colorectal cancer may be related to the regulation of immune inflammatory response,autophagy,cell proliferation and apoptosis,and has the characteristics of multi-target,multi-path and multi-system mechanism.

Key words: Canmei Decoction, colorectal cancer, network pharmacology, mechanism of action, quercetin

中图分类号:  R961.1