整合网络药理学和体外实验探究夏枯草-半枝莲药对治疗乳腺癌的作用及机制  

The effect and mechanism of Prunella vulgaris-Scutellaria barbata herb pairs in the treatment of breast cancer by integrating network pharmacology and in vitro experiments

作　　者：刘苏 陈洪晓 金乐 张慧慧 张蕾[2] 陈昭琳[1,2] Liu Su;Chen Hongxiao;Jin Le;Zhang Huihui;Zhang Lei;Chen Zhaolin(School of Pharmacy,Anhui Medical University,Hefei 230032;Dept of Pharmacy,The First Affiliated Hospital of USTC(Anhui Provincial Hospital),Hefei 230001;Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-Inflammatory and Immune Medicine,Hefei 230032)

机构地区：[1]安徽医科大学药学院,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,合肥230001 [3]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,合肥230032

出　　处：《安徽医科大学学报》2024年第7期1134-1142,共9页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:2208085MH252、2108085 MH311);安徽省高校优秀青年科研项目(编号:2023AH030114)。

摘　　要：目的采用网络药理学和体外细胞实验探究夏枯草-半枝莲药对抗乳腺癌的作用机制。方法运用中药系统药理学数据库与分析平台筛选出夏枯草-半枝莲药对的有效成分和靶点;利用GeneCards、OMIM数据库找到乳腺癌靶点,进而构建药物-活性成分-关键靶点网络和蛋白-蛋白相互作用(PPI)网络;运用R语言进行GO功能和KEGG通路富集分析以及生存分析;将筛选出的活性成分和核心靶点进行分子对接验证;采用CCK-8法检测细胞活力;EdU、流式实验检测细胞增殖和凋亡情况;Western blot法检测p-AKT1、AKT1、β-catenin和c-MYC的蛋白表达水平。结果通过数据分析共筛选出有效成分36个,交集靶点105个,其核心成分是槲皮素、木犀草素、山奈酚、汉黄芩素、黄芩素;通过PPI和生存分析得出其关键靶点是AKT1、ESR1、CASP3、MYC;GO分析共包含4303条富集结果,KEGG分析共包含232条通路;分子对接显示核心成分与关键靶点均具有较强的结合能力。细胞实验证明,核心活性成分槲皮素可以抑制乳腺癌细胞的增殖并促进其凋亡(P<0.05),下调p-AKT1、β-catenin和c-MYC蛋白的表达水平(P<0.05)。结论夏枯草-半枝莲药对中活性成分槲皮素可能通过AKT1/β-catenin信号通路来发挥作用,为其治疗乳腺癌的作用机制研究提供了科学参考。Objective To explore the mechanism of Prunella vulgaris and Scutellaria barbata herb pair against breast cancer based on network pharmacology and in vitro cell experiments.Methods The effective components and targets of Prunella vulgaris and Scutellaria barbata herb pair were screened.GeneCards and OMIM databases were used to find breast cancer targets,and then drug-active ingredient-key target network and protein-protein interaction(PPI)were constructed.R language was used to perform GO function and KEGG pathway enrichment analysis and survival analysis.Then the screened active components and core targets were verified by molecular docking.Cell viability was detected by CCK-8 assay.EdU and flow cytometry were used to detect cell proliferation and apoptosis.The protein expression levels of p-AKT1,AKT1,β-catenin and c-MYC were detected by Western blot.Results Through databases analysis,a total of 36 active components and 105 intersection targets were screened out,the core components were quercetin,luteolin,kaempferol,wogonin and baicalein.Through PPI and survival analysis,the key targets were AKT1,ESR1,CASP3 and MYC.GO analysis contained 4303 enrichment results,KEGG analysis contained 232 pathways.Molecular docking showed that the core components had strong binding ability with the key targets.Cell experiments showed that the core active ingredient quercetin could inhibit the proliferation of breast cancer cells and promote their apoptosis(P<0.05),and down-regulate the expression levels of p-AKT1,β-catenin and c-MYC proteins(P<0.05).Conclusion The active components quercetin in Prunella vulgaris and Scutellaria barbata herb pair may play a role through AKT1/β-catenin signaling pathway,which provides a scientific reference for the study of its mechanism of action in the treatment of breast cancer.

关 键 词：网络药理学 夏枯草 半枝莲 分子对接 细胞实验 作用机制 

分 类 号：R966[医药卫生—药理学]