雪菊总黄酮对结肠癌细胞抗瘤作用的网络药理学机制研究  

Mechanisms of chrysanthemum total flavonoids on anti-tumor effect of colon cancer cells based on network pharmacology

作　　者：伊丽米热·吾甫尔 库热西·玉努斯 国鲁源 卡斯木江·阿西木江 吴桂霞[1] ZHONG Li Yilimire Wufuer;Kurexi Yunusi;GUO Luyuan;ZHONG Li;Kasimujiang Aximujiang;WU Guixia(School of Basic Medical Sciences,2Uyghur Medical College,Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学基础医学院,乌鲁木齐830011 [2]新疆医科大学维吾尔医学院,乌鲁木齐830011 [3]School of Basic Medical Sciences,Xinjiang Medical University,Urumqi 830011,China

出　　处：《新疆医科大学学报》2021年第6期670-678,共9页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区自然科学基金(2019D01C218)。

摘　　要：目的基于网络药理学和体外细胞实验,探讨雪菊总黄酮对人结肠腺癌Caco2细胞抗瘤作用的主要活性成分及潜在的作用机制。方法利用TCMSP、STRING、DisGeNET、OmicShare、Cytoscape数据库及平台筛选雪菊总黄酮有效活性成分及其对结肠癌抗瘤的潜在靶点,通过生物学信息注释数据库(DAVID)对预测靶点进行基因本体(GO)和京都基因与基因组百科全书(KEGG)通路富集分析,构建“雪菊总黄酮-靶点-通路”网络。利用Auto dock vina进行分子对接,分析雪菊总黄酮化合物与关键靶点的结合活性。基于网络药理学的分析结果,通过雪菊总黄酮干预Caco2的体外实验进行核心靶点和通路的初步验证。结果筛选出雪菊总黄酮的12个活性成分及对结肠癌的63个抗瘤靶点,KEGG通路富集分析结果显示,抗瘤作用主要被富集在与细胞增殖、凋亡和迁移调控等相关的PI3K-Akt等15个与肿瘤相关的信号通路。分子对接提示雪菊总黄酮活性成分表没食子儿茶素没食子酸酯、黄芩素、金合欢素及黄豆黄素与AKT1,Bcl-2、Caspase-3和MMP-9等关键靶点结合性较好。体外细胞实验结果表明,雪菊总黄酮明显抑制了Caco2细胞的增殖、促进细胞凋亡、抑制细胞迁移。Western blot的结果表明雪菊总黄酮上调Bax/Bcl-2的比值、下调了Caspase-3和MMP-9的表达。结论雪菊总黄酮可通过多靶点、多途径实现干预结肠癌的作用,其抗肿瘤活性可能与PI3K-Akt信号通路中Bax、Bcl-2、Caspase-3和MMP-9的表达调节有关。Objective To explore the mechanism of total flavonoids of chrysanthemum against human colonic adenocarcinoma cells line Caco2 based on network pharmacological and cell experiments in vitro.Methods Using the TCMSP,STRING,DisGeNET,OmicShare,Cytoscape database and platform to screen the active components of total flavonoids and their potential targets against colon cancer,to predict targets through GO and KEGG pathway enrichment analysis by DAVID,and to construct"total flavonoids of chrysanthemum-target-pathway"network.Conducting molecular docking by Auto dock vina to analyze the binding activity of total flavonoid compounds and key targets.And then to preliminarily verify the prediction by cell experiments of total flavonoids.Results The results showed that 12 active components of total flavonoids of chrysanthemum and 63 anti-tumor targets for colon cancer were screened.KEGG analysis showed that anti-tumor effect was mainly enriched in 15 tumor-related signaling pathways,such as PI3KAkt related to cell proliferation,apoptosis and migration regulation.Molecular docking suggests that the active components of total flavonoids:(-)-Epigallocatechin_gallate,baicalein,acacetin and astilbin combinded well with the key targets such as AKT1,Bcl-2,Caspase-3 and MMP-9.The results of the cell experiment showed the total flavonoids of chrysanthemum significantly inhibited the proliferation of Caco2 cells,promoted apoptosis and inhibited cell migration.Western blotting results showed that the total flavonoids upregulated the ratio of Bax/Bcl-2 and downregulated the expression of Caspase-3 and MMP-9.Conclusion The total flavonoids of chrysanthemum can intervene in colon cancer through multiple targets and pathways.Its anti-tumor activity may be related to the regulation of Bax,Bcl-2,Caspase-3 and MMP-9 expression in PI3K-Akt signaling pathway.

关 键 词：网络药理学 雪菊总黄酮 结肠癌 凋亡 

分 类 号：R735.3[医药卫生—肿瘤]