基于网络药理学与分子对接及实验验证探讨苦杏仁-桔梗药对治疗肺癌的作用机制  

Exploring the Mechanism of Action of Armeniacae Semen Amarum-Platycodonis Radix in the Treatment of Lung Cancer Based on Network Pharmacology and Molecular Docking and Experimental Validation

作　　者：刘远财 周夏成 陈卓[2] 杨佳颖 张爱琴[2] LIU Yuancai;ZHOU Xiacheng;CHEN Zhuo;YANG Jiaying;ZHANG Aiqin

机构地区：[1]浙江中医药大学第二临床医学院,浙江杭州310053 [2]浙江省肿瘤医院,浙江杭州310022

出　　处：《新中医》2023年第11期146-155,共10页New Chinese Medicine

基　　金：浙江省中医药科技计划重点项目(2021ZZ008)。

摘　　要：目的:运用网络药理学方法及分子对接初步预测苦杏仁-桔梗药对治疗肺癌的主要靶点及其作用机制,再通过体外实验验证部分关键靶点及其通路。方法:通过中药系统药理学数据库与分析平台(TCMSP)、SwissTargetPrediction、GeneCard数据库获取活性成分和靶点,利用UniProt数据库将靶点标准化,通过在线veny2.1绘制韦恩图。运用STRING数据库绘制靶点的PPI网络图,使用Cytoscape3.8.2软件建立“药物-成分-靶点”“核心蛋白-蛋白”网络关系图,利用Matescape数据库进行GO和KEGG通路富集分析。通过PyMOL、AutoDockTool 1.5.6软件进行分子对接。用不同浓度的小鼠含药血清培养Lewis肺癌细胞,CCK8检测细胞活性,流式细胞Annen V-FITC/PI标记法检测细胞凋亡率,实时荧光定量PCR技术、Western Blot检测Lewis肺癌细胞中SRC蛋白表达水平。结果:获得苦杏仁-桔梗有效活性成分23个,靶点为227个,关键靶点为SRC,KEGG富集程度较高的为PI3K-Akt信号通路、MAPK信号通路等。分子对接显示结合能良好。体外实验显示苦杏仁-桔梗药对可抑制Lewis肺癌细胞增殖,诱导其凋亡,并能下调SRC蛋白表达水平。结论:苦杏仁-桔梗药对能抑制SRC的表达,可能通过调控PD-1和PD-L1通路、Ras信号通路、PI3K-Akt信号通路、MAPK信号通路降低肿瘤细胞的侵袭、迁移、增殖、分化等能力,增强机体免疫力,从而治疗肺癌。Objective:To preliminarily predict the main targets and mechanisms of Armeniacae Semen Amarum-Platycodonis Radix in the treatment of lung cancer using network pharmacological methods and molecular docking,and then verify some key targets and pathways through in vitro experiments.Methods:Active ingredients and targets were obtained through the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform(TCMSP),SwissTargetPrediction and GeneCard databases.The targets were standardized using the UniProt database,and the Venn diagram was drawn through online veny2.1.Using STRING database to draw the target PPI network diagram,using Cytascape3.8.2 software to establish the"drug-component-target"and"core protein-protein"network relationship diagram,and using Matescape database to conduct GO and KEGG pathway enrichment analysis.Molecular docking is performed through PyMOL and AutoDockTool 1.5.6 software.Lewis lung cancer cell was cultured with different concentrations of mouse drug-containing serum.Cell activity was detected by CCK8,apoptosis rate was detected by flow cytometry using Annen V-FITC/PI labeling,and the expression level of SRC protein in Lewis lung cancer cell was detected by real-time fluorescence quantitative PCR and Western Blot.Results:A total of 23 effective active components were obtained from Armeniacae Semen Amarum-Platycodonis Radix,with 227 target.The key target was SRC,and the PI3K-Akt signaling pathway and MAPK signaling pathway with higher KEGG enrichment were found.Molecular docking showed good docking performance.In vitro experiments showed that the Armeniacae Semen Amarum-Platycodonis Radix drug pair could inhibit the proliferation of Lewis lung cancer cell,induce the apoptosis,and down-regulate the expression level of SRC protein.Conclusion:The Armeniacae Semen Amarum-Platycodonis Radix drug pair can inhibit the expression of SRC,possibly by regulating the PD-1 and PD-L1 pathways,Ras signaling pathway,PI3K-Akt signaling pathway,and MAPK signaling pathway to reduce the inv

关 键 词：肺癌 苦杏仁-桔梗 SRC 网络药理 体外实验 

分 类 号：R734.2[医药卫生—肿瘤]