小檗碱抑制乳腺癌细胞增殖的网络药理学分析及体外细胞验证  

Network pharmacological analysis of berberine inhibiting breast cancer cell proliferation and in vitro cell validation

作　　者：张慧慧 金乐 刘苏 陈洪晓 陈昭琳 唐丽琴[1,2] ZHANG Huihui;JIN Le;LIU Su;CHEN Hongxiao;CHEN Zhaolin;TANG Liqin(Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-Inflammatory and Immune Medicine,Hefei 230032,Anhui,China;Department of Pharmacy,The First Affiliated Hospital of USTC,Division of Life Sciences and Medicine,University of Science and Technology of China,Hefei 230001,Anhui,China)

机构地区：[1]安徽医科大学临床药理研究所,抗炎免疫药物教育部重点实验室,安徽合肥230032 [2]中国科学技术大学附属第一医院(安徽省立医院)药学部,安徽合肥230001

出　　处：《中国临床药理学与治疗学》2025年第3期332-338,共7页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金面上项目(82174031);安徽省自然科学基金(2208085MH252);安徽省高校优秀青年科研(2023AH030114)。

摘　　要：目的:基于网络药理学和体外细胞实验探究小檗碱对乳腺癌细胞的作用机制。方法:首先,以小檗碱与乳腺癌作为研究对象,通过VEEN图筛选两者交集靶点,利用R语言进行GO功能和KEGG富集分析,利用AutodockVina和Pymol软件进行分子对接及可视化。然后,小檗碱处理乳腺癌MCF-7细胞24 h后,进行体外细胞实验验证。采用CCK-8法检测细胞活力,Edu和平板克隆检测细胞增殖克隆能力,An-nexinV-FITC/PI双染法和Westernblot检测细胞凋亡。利用激光共聚焦和CETSA验证小檗碱和AKT1蛋白的结合作用。结果:网络药理学结果表明,小檗碱与核心靶点AKT1、AKT2、MAPK3等均有较好的结合。小檗碱(20、40、80μmol/L)能明显抑制MCF-7细胞的增殖克隆能力并且呈浓度依赖性(P<0.05,P<0.01)。激光共聚焦和CETSA实验结果显示,小檗碱和AKT1具有结合作用,且两者结合之后稳定性增强。结论:小檗碱通过与AKT1蛋白靶向结合,抑制人乳腺癌MCF-7细胞增殖并诱导其凋亡。AIM:To explore the mechanism of berberine on breast cancer cells based on network pharmacology and in vitro cell experiments.METHODS:Firstly,berberine and breast cancer were taken as the research objects,the intersection targets of the two were screened by VEEN diagram,GO function and KEGG enrichment analysis were performed by R language,and molecular docking and visualization were carried out by Autodock Vina and Pymol software.Then,berberine treated breast cancer MCF-7 cells for 24 h,and then in vitro cell experiments were performed.CCK-8 was used to detect cell viability,Edu and plate cloning were used to detect cell proliferation and cloning,and apoptosis was detected by An-nexin V-FITC/PI double staining and Western blot.Laser confocal and CETSA were used to verify the binding effect of berberine and AKT1 protein.RESULTS:The results of network pharmacology showed that berberine had a good binding to the core targets AKT1,AKT2 and MAPK3.Berberine(20,40,80μmol/L)significantly inhibited the proliferation and cloning ability of MCF-7 cells in a concentration-dependent manner(P<0.05,P<0.01).The results of laser confocal and CETSA experiments showed that berberine and AKT1 had a binding effect,and the stability of the two was enhanced after the combination.CONCLUSION:Berberine inhibits MCF-7 cell proliferation and induces apoptosis in human breast cancer cells by targeting binding to AKT1 protein.

关 键 词：网络药理学 小檗碱 乳腺癌 AKT1 

分 类 号：R965.2[医药卫生—药理学]