摘要
目的采用网络药理学探讨黄芪-丹参治疗肝纤维化的潜在作用机制,并通过小鼠体内实验进行验证。方法利用中药系统药理学数据库和疾病靶点数据库筛选黄芪-丹参的主要活性成分及其作用靶点,并进行基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)富集分析,选择磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)通路,利用Cytoscape3.7.2软件构建黄芪-丹参的核心成分-关键靶点-目标通路的网络图。将50只SPF级KM小鼠,随机分成正常对照组,模型组,黄芪-丹参低、中、高剂量(2.5、5.0和10.0 g·kg^(-1)·d^(-1),黄芪丹参生药质量比11)组,每组10只。检测实验处理前后小鼠体重、肝重、肝脏组织病理以及关键靶蛋白和mRNA表达水平变化。结果黄芪-丹参活性靶点与肝纤维化疾病靶点映射得到183个共有靶点。通过“黄芪丹参核心成分-关键靶点-目标通路”网络分析,血管内皮生长因子α(VEGFA)、碱性成纤维细胞生长因子2(FGF2)、表皮生长因子受体(EGFR)、有丝分裂原活化蛋白激酶1(MAPK1)和糖原合成酶激酶3β(GSK-3β)为关键靶点。与模型组相比,黄芪-丹参各剂量组肝纤维化情况均有缓解,EGFR、MAPK1、VEGFA和GSK-3β的mRNA表达水平均显著降低(P<0.05),FGF2的mRNA表达水平显著升高(P<0.05);高剂量组的AKT、p-PI3K,中剂量组的p-AKT、PI3K,低剂量组的p-AKT和p-PI3K蛋白表达水平显著升高(P<0.05)。结论黄芪-丹参的核心成分可能通过调控PI3K/AKT/mTOR通路作用于VEGFA、FFG2、EGFR、MAPK1和GSK-3β而改善肝纤维化。
AIM To explore the mechanism of Astragalus membranaceus-Salvia miltiorrhiza in the treatment of liver fibrosis using network pharmacology and verified in vivo experiments in mice.METHODS The main active ingredients of Astragalus membranaceus-Salvia miltiorrhiza were screened and the main active targets against liver fibrosis were screened by network pharmacology.The Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were conducted.The phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/rapamycin target protein(mTOR)pathway was selected,and a network diagram of the core component,key target,and target pathways of Astragalus membranaceus-Salvia miltiorrhiza was constructed using Cytoscape 3.7.2 software.Fifty KM mice were randomly divided into control group,model group,and low,medium,and high dose(25,50 and 100 mg·kg^(-1),Astragalus membranaceus-Salvia miltiorrhiza raw drug mass ratio 11)groups,10 mice in each group.The changes in body weight,liver weight,liver histopathology,and expression level of key target protein and mRNA in mice before and after the treatment were detected.RESULTS A total of 183 common targets were obtained by mapping the active targets of Astragalus membranaceus-Salvia miltiorrhiza and disease targets of liver fibrosis.Through the analysis of the network diagram of Astragalus membranaceus-Salvia miltiorrhiza core ingredients-key targets-target pathways,vascular endothelial growth factor alpha(VEGFA),basic fibroblast growth factor 2(FGF2),epidermal growth factor receptor(EGFR),mitogen activated protein kinase 1(MAPK1)and glycogen synthase kinase-3 beta(GSK-3β)are key targets.Compared with the model group,the liver fibrosis were effectively alleviated,the expression levels of EGFR,MAPK1,VEGFA and GSK-3βmRNA were significantly decreased(P<0.05),and FGF2 mRNA expression level was obviously increased(P<0.05)in Astragalus membranaceus-Salvia miltiorrhiza groups.The expression levels of AKT,p-PI3K in the high-dose group,p-AKT,PI3K in the medium-dose group,p-AKT and p-PI3K protein in the low-dose groups were significantly increased(P<0.05).CONCLUSION The core ingredients of Astragalus membranaceus-Salvia miltiorrhiza may improve liver fibrosis to act on VEGFA,FGF2,EGFR,MAPK1 and GSK-3βby regulating the PI3K/AKT/mTOR pathway.
作者
周佳林
饶颖
刘翔
李伟阁
李文杰
戴琦
ZHOU Jia-lin;RAO Ying;LIU Xiang;LI Wei-ge;LI Wen-jie;DAI Qi(Jiangxi University of Traditional Chinese Medicine,Nanchang JIANGXI 330000,China;The Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine,Nanchang JIANGXI 330000,China)
出处
《中国新药与临床杂志》
CAS
CSCD
北大核心
2023年第11期750-759,共10页
Chinese Journal of New Drugs and Clinical Remedies
基金
国家自然科学基金(81860791,82160903)
江西省中医药科技计划项目(2020A005)
江西省自然科学基金青年基金项目(20122BAB216060)。
关键词
黄芪
丹参
肝纤维化
网络药理学
动物实验
药理作用分子作用机制
Astragalus membranaceus
Salvia miltiorrhiza
hepatic fibrosis
network pharmacology
animal experimentation
molecular mechanisms of pharmacological action
