基于网络药理学及实验研究探究黄芪-莪术药对治疗胃癌的作用机制  

Mechanism of Astragali Radix-Curcumae Rhizoma in treating gastric cancer based on network pharmacology and experimental verification

作　　者：谭喜莹 陶靖[2] 张宇[3] 顾茹辛 TAN Xi-ying;TAO Jing;ZHANG Yu;GU Ru-xin(Department of Pharmacy,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;China Pharmaceutical University,Nanjing 210009,China;Nanjing Medical University,Nanjing 211166,China)

机构地区：[1]南京中医药大学附属医院药学部,江苏南京210029 [2]中国药科大学,江苏南京210009 [3]南京医科大学,江苏南京211166

出　　处：《中国中药杂志》2023年第18期5056-5067,共12页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82003961);江苏省中医药科技发展计划项目(YB201921);北京医卫健康公益基金会项目(YWKYQ4007)。

摘　　要：该研究目的在于利用网络药理学方法探究黄芪-莪术药对治疗胃癌的作用机制,并通过胃癌细胞SGC7901体外模型对黄芪-莪术的药效和关键靶点进行实验验证。首先,采用CCK-8法证明黄芪-莪术对胃癌细胞SGC7901增殖的直接影响。然后,使用网络药理学的方法探究黄芪-莪术药对治疗胃癌的关键活性成分、关键靶点和关键信号通路。结果显示,黄芪-莪术药对包含18个潜在活性成分,例如槲皮素、山柰酚和姜黄素等。基因本体论功能富集及京都基因和基因组百科全书通路富集分析提示,黄芪-莪术药对治疗胃癌的主要靶点涉及蛋白丝氨酸/苏氨酸激酶活性的调节、MAPK活性的激活、半胱氨酸型内肽酶活性和蛋白丝氨酸/苏氨酸激酶活性的负调节等。HIF-1信号通路、ABC转运蛋白、细胞色素P450对异生素的代谢、p53信号通路和细胞凋亡等信号通路是黄芪-莪术作用于胃癌的关键通路。随后,在体外实验中证明,黄芪-莪术药对能够显著下调多药耐药基因(ABCB1)、表皮生长因子受体(EGFR)、磷酸化蛋白激酶B(p-AKT)、低氧诱导因子-1(HIF1A)、B-细胞淋巴瘤因子2(BCL2)、乳腺癌易感基因1(BRCA1)、DNA聚合酶θ(POLH)、核苷酸还原酶M1(RRM1)、切除修复交叉互补基因1(ERCC1)的表达,并上调肿瘤蛋白p53(TP53)和半胱氨酸天冬氨酸蛋白酶3(CAPS3)表达。最后,利用癌症基因组图谱计划(TCGA)数据库中的胃癌患者基因表达和临床信息数据,应用多变量COX回归模型证明了黄芪-莪术作用的关键靶点与胃癌患者不良预后的相关性,并通过LASSO算法进行特征选择。结果显示,EGFR、HIF1A、TP53、POLH、RRM1和ERCC1与胃癌患者的生存期密切相关。黄芪-莪术对上述多个靶点及通路的作用,通过影响胃癌细胞的DNA修复、生存和凋亡等相关基因的表达,从而在胃癌辅助治疗中发挥重要的作用。This study aims to investigate the mechanism of Astragali Radix-Curcumae Rhizoma(HQEZ)in the treatment of gastric cancer based on network pharmacology.Further,the SGC7901 cell model of gast ric cancer was employed to validate the efficacy and key targets of the herb pair.Firstly,the CCK8 assay was employed to evaluate the direct effect of HQEZ on the proliferation of gastric cancer SGC7901 cells.Then,network pharmacology methods were empl oyed to investigate the active ingredients,key targets,and key signaling pathways involved in the treatment of gastric cancer with HQEZ.The result s showed that HQEZ contained 18 potential active ingredients,such as quercetin,naringenin,and curcumin.The results of gene ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment suggested that the main targets of HQEZ in treating gastric cancer were involved in the regulation of protein serine/threonine kin ase activity,activation of mitogen-activated protein kinase(MAPK)activity,cysteine-type endopeptidase activity,and negative regulation of protein serine/threonine kinase activity.The hypoxia-inducible factor-1(HIF-1)signaling pathway,ATP-binding cassette(ABC)transporters,cytochrome P450-mediated metabolism of xenobiotics,p53 signaling pathway,and cell apoptosis were key signaling pathways of HQEZ in treating gastric cancer.The cell experiments demonstrated that HQEZ significantly downregulated the expression of ATP-binding cassette subfamily B member 1(ABCB1),epidermal growth factor receptor(EGFR),AKT serine/threonine kinase 1(AKT1),HIF1A,B-cell lymphoma 2(BCL2),breast cancer 1(BRCA1),DNA polymerase theta(POLH),ribonucleotide reductase M1(RRM1),and excision repair cross-complementation group 1(ERCC1),and upregulated the expression of TP53 and cysteinyl aspartate-specific proteinase(CAPS3).Finally,a multivariate COX regression model was adopted to study the relationship between gene expression and clinical information data of gastric cancer patients in the TCGA database,which demonstrat

关 键 词：胃癌 黄芪-莪术 药对 网络药理学 作用机制 

分 类 号：R285[医药卫生—中药学]