摘要
目的基于网络药理学、分子对接及动物实验验证揭示复方黄芪汤抗肺癌分子机制。方法通过网络药理学预测复方黄芪汤抗肺癌的核心成分、靶基因和关键信号通路并进行分子对接;构建Lewis荷瘤小鼠模型,设立空白对照组、荷瘤对照组、Pd^(-1)抗体组(10 mg·kg^(-1))及复方黄芪汤低、高剂量组(30、60 g·kg^(-1)),统计肿瘤质量和抑瘤率;苏木精-伊红(HE)染色法检测肿瘤及肺组织病理变化,免疫组化法检测肿瘤组织中微血管密度蛋白(FactorⅧ)表达,酶联免疫吸附测定法(ELISA)检测血清中血管内皮生长因子(VEGF)含量,Western Blot法检测肿瘤组织中磷酸化蛋白激酶B(p-AKT)和蛋白激酶B(AKT)蛋白表达。结果从数据库筛选出复方黄芪汤活性成分56个,交集靶点181个,核心靶点9个,对应活性成分8个;基因本体(GO)分析及基因组百科全书(KEGG)信号通路富集分析结果显示,其主要通过调控PI3K-AKT、JAK-STAT、EGFR酪氨酸激酶抑制剂耐药等信号通路抗肺癌。分子对接提示黄芪紫檀烷苷与VEGF及山柰酚、熊竹素、异鼠李素与蛋白激酶Bα(AKT1)具有良好结合力。动物实验验证结果显示,与荷瘤对照组比较,复方黄芪汤高剂量组能显著抑制肿瘤生长,降低肿瘤质量(P<0.01)。HE染色结果显示肿瘤细胞的细胞核大且核分裂相明显,而复方黄芪汤干预后肿瘤细胞排列松散,细胞间隙大,细胞活跃度低。与荷瘤对照组比较,复方黄芪汤高剂量组能显著降低FactorⅧ水平(P<0.001)及VEGF含量(P<0.01);Western Blot结果显示其能显著下调p-AKT/AKT比值(P<0.01)。结论复方黄芪汤可能通过干预VEGF-AKT信号通路,抑制肿瘤血管生成,进而发挥抑瘤作用。
Objective To elucidate the molecular mechanism of Compound Astragali Radix Decoction(CARD)against lung cancer through network pharmacology,molecular docking,and experimental validation in animals.Methods Network pharmacology was employed to predict the core components,target genes,and key signaling pathways of CARD against lung cancer,followed by molecular docking verification.A Lewis tumor-bearing mouse model was established and divided into blank control,tumor-bearing control,PD-1 antibody(10 mg·kg^(-1)),low-dose CARD(30 g·kg^(-1)),and high-dose CARD(60 g·kg^(-1))groups.Tumor mass and inhibition rate were measured.Hematoxylin-eosin(HE)staining was used to examine pathological changes in tumor and lung tissues;immunohistochemistry was performed to detect microvessel density(FactorⅧ)expression in tumor tissues;enzyme-linked immunosorbent assay(ELISA)was used to measure serum vascular endothelial growth factor(VEGF)levels;and Western Blot was employed to assess phosphorylated protein kinase B(p-AKT)and total AKT protein expression in tumor tissues.Results There were 56 active components and 181 common targets of CARD were identified from databases,with 9 core targets and 8 corresponding active components selected.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses indicated that CARD exerts anti-lung cancer effects primarily by regulating the PI3K-AKT,JAK-STAT,and EGFR tyrosine kinase inhibitor resistance signaling pathways.Molecular docking demonstrated strong binding affinity between astrapterocarpan and VEGF,as well as between kaempferol,jaranol,and isorhamnetin with protein kinase Bα(AKT1).Animal experiments revealed that,compared with the tumor-bearing control group,the high-dose CARD group significantly inhibited tumor growth and reduced tumor mass(P<0.01).HE staining showed that tumor cells in the control group had large nuclei and prominent mitotic figures,whereas CARD intervention resulted in loosely arranged tumor cells with wide intercellular spaces and reduced cell activity.The high-dose CARD group significantly decreased FactorⅧlevels(P<0.001)and VEGF content(P<0.01)compared with the tumor-bearing control group.Western Blot results indicated a significant downregulation of the p-AKT/AKT ratio(P<0.01).Conclusion Compound Astragali Radix Decoction may inhibit tumor angiogenesis and exert anti-tumor effects by interfering with the VEGF-AKT signaling pathway.
作者
钟怡
尹春燕
廉源沛
蔡佳丽
王地均
聂雪珂
刘产明
申春悌
颜晓静
ZHONG Yi;YIN Chunyan;LIAN Yuanpei;CAI Jiali;WANG Dijun;NIE Xueke;LIU Chanming;SHEN Chunti;YAN Xiaojing(Changzhou Hospital Affiliated to Nanjing University of Chinese Medicine,Changzhou Key Laboratory of Human Use Experiment Research&Transformation of Menghe Medical School,Changzhou 213003 Jiangsu,China)
出处
《中药新药与临床药理》
北大核心
2026年第1期70-81,共12页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金项目(82074016,81603296)
江苏省中医药管理局项目(2020ZX18)
常州市卫生健康委员会重大项目(ZD202028)
常州市科技计划项目(CM20223006,CJ20219008)
2024年江苏省研究生科研与实践创新计划项目(SJCX24_0919)。
关键词
复方黄芪汤
肺癌
抑瘤网络药理学
VEGF-AKT信号通路
血管新生
实验验证
小鼠
Compound Astragali Radix Decoction
lung cancer
anti-tumor effects network pharmacology
VEGFAKT signaling pathway
angiogenesis
experimental validation
mice
