基于网络药理学探讨平消胶囊通过AKT1/β-catenin信号促进乳腺癌凋亡的作用机制

Mechanism of Pingxiao capsule promoting breast cancer apoptosis through AKT1/β-catenin signal based on network pharmacology

目的:探究平消胶囊治疗乳腺癌的潜在作用机制。方法:利用TCMSP、TCM-ID、GeneCards等数据库筛选平消胶囊与乳腺癌的相关靶点;Cytoscape软件构建“药物-靶点-疾病”网络;运用R软件进行GO和KEGG分析;使用Autodock Vina和Pymol软件进行平消胶囊活性成分与核心靶点的分子对接及可视化;通过R软件survival包对核心靶点进行分析,筛选出与总体生存时间密切相关的基因;通过CCK-8法检测细胞活力;流式细胞术检测细胞凋亡;Western blot检测p-AKT1、β-catenin和cyclinD1的蛋白表达水平。结果:筛选出药物靶点194个,疾病靶点1612个,韦恩图求交集靶点共127个,核心靶点主要包括TB53、AKT1、TNF、CASP3等20个;GO分析主要与氧化应激反应、细胞对化学反应的调控等生物活性有关;KEGG分析主要通过PI3K-AKT信号通路、TNF信号通路、IL-17信号通路等通路有关;分子对接结果显示活性成分与核心靶点AKT1、MAPK1、RELA均有较好的结合;细胞实验表明槲皮素(40、80、120μmol/L)促进乳腺癌细胞凋亡;Western blot检测显示随着不同浓度的槲皮素处理乳腺癌细胞48 h后,p-AKT1、β-catenin和cyclinD1的蛋白表达量均不同程度地降低。结论:网络药理学与细胞实验证实平消胶囊可能通过调节AKT1/β-catenin信号通路,发挥其抗乳腺癌作用。

AIM:To explore the potential mecha-nism of action of Pingxiao capsule in the treatment of breast cancer.METHODS:TCMSP,TCM-ID,Gene-Cards and other databases were used to screen the related targets of Pingxiao capsule and breast can-cer.Cytoscape software builds drug-target-disease networks.R software was used for GO and KEGG analysis.Autodock Vina and Pymol software were used for molecular docking and visualization of Pingxiao capsule active ingredients and core tar-docking results showed that the active constituents were well combined with the core targets AKT1,MAPK1 and RELA.Cell experiments showed that quercetin(40,80,120μmol/L)promoted apoptosis of breast cancer cells.Western blot analysis showed that the protein expressions of p-AKT1,β-catenin and cyclinD1 decreased with different con-centrations of quercetin treated for 48h.CONCLU-SION:Network pharmacology and cell experiments confirmed that Pingxiao capsule may exert its anti-breast cancer effect by regulating AKT1/β-catenin signaling pathway.