基于网络药理学和实验验证探讨光甘草定治疗去势抵抗性前列腺癌的作用机制  

Molecular mechanism of glabridin in the treatment of castration-resistant prostate cancer based on network pharmacology and experimental verification

作　　者：谭武宾 周松 康海 蒋湘勇 毛正 杨科[1] 李铁求 TAN Wu-bin;ZHOU Song;KANG Hai;JIANG Xiang-yong;MAO Zheng;YANG Ke;LI Tie-qiu(Hunan Provincial People′s Hospital(The First Affiliated Hospital of Hunan Normal University),Changsha 410002,China;People′s Hospital of Changsha County(Hunan Provincial People′s Hospital Xingsha Branch),Changsha 410100,China)

机构地区：[1]湖南省人民医院(湖南师范大学附属第一医院),长沙410002 [2]长沙县人民医院(湖南省人民医院星沙院区),长沙410100

出　　处：《天然产物研究与开发》2023年第2期310-318,259,共10页Natural Product Research and Development

基　　金：湖南省中医药管理局项目(2021041);国家重点临床专科建设项目(2022-2);长沙市科技局项目(kq2208112)。

摘　　要：本研究结合网络药理学、分子对接和体外细胞实验探讨光甘草定治疗去势抵抗性前列腺癌的作用机制。利用中药数据库HERB、TCMIO、TCMSP、PharmMapper和BATMAN-TCM、Swiss Target Prediction数据库共筛选出248个药物靶点,利用DisGeNET数据库收集到683个疾病靶点,通过韦恩图工具获取药物-疾病靶点55个。通过PPI分析以及拓扑学分析得到AKT1、TP53、ESR1、EGFR等10个核心靶点蛋白。使用R studio软件的“clusterProfiler”包进行GO功能富集分析和KEGG通路富集分析,GO富集分析分析得到847个条目(P<0.01),涉及对类固醇激素的反应、蛋白激酶B信号、对营养水平的反应等,KEGG富集分析得到PI3K/AKT信号通路、蛋白聚糖在癌症中的作用、前列腺癌等105个相关信号通路(P<0.01)。运用AutoDock软件进行分子对接,结果表明光甘草定与核心靶点具有较好的结合性。CCK8法检测细胞增殖发现光甘草定可抑制PC-3细胞增殖(P<0.05),流式细胞术和DAPI染色镜下观察发现光甘草定可促进细胞凋亡(P<0.05),Western blot实验发现光甘草定可抑制AKT的磷酸化(P<0.05)。综上,初步研究发现光甘草定可能通过调控AKT1、TP53、ESR1、EGFR等核心蛋白以及PI3K/AKT等信号通路发挥抗去势抵抗性前列腺癌的作用。To investigate the mechanism of action of glabridin in the treatment of castration-resistant prostate cancer(CRPC),we combined network pharmacology,molecular docking and in vitro cellular assays.A total of 248 drug targets were screened by HERB,TCMIO,TCMSP,PharmMapper and Batmant-TCM,Swiss Target Prediction database,and 683 disease targets were collected by DisGeNET database.Fifty-five drug-disease targets were obtained through the Venn diagram tool.Through PPI analysis and topological analysis,ten core target proteins such as AKT1,TP53,ESR1 and EGFR were obtained.The"clusterProfiler"package of R Studio software was used for GO functional enrichment analysis and KEGG pathway enrichment analysis.These targets are involved 847 GO items(P<0.01),involving response to steroid hormone,protein kinase B signaling,response to nutrient levels,etc.KEGG enrichment analysis revealed 105 signaling pathways related to PI3K/AKT signaling pathway,Proteoglycans in cancer,prostate cancer and other signaling pathways(P<0.01).The results of molecular docking using AutoDock software showed that glabridin had good binding ability with the core target.CCK8 assay showed that glabridin could inhibit PC-3 cell proliferation(P<0.01),flow cytometry and DAPI staining showed that glabridin could promote cell apoptosis(P<0.01),and Western blot showed that glabridin could inhibit the phosphorylation of AKT(P<0.01).In conclusion,our initial findings show that Glabridin may regulate AKT1,TP53,ESR1,EGFR and other proteins and through some important signaling pathways such as PI3K/AKT signaling pathway exerted anti-castration-resistant prostate cancer effects.

关 键 词：光甘草定 去势抵抗性前列腺癌 网络药理学 分子对接 

分 类 号：R285[医药卫生—中药学] R917[医药卫生—中医学]