基于网络药理学和动物实验研究HIF-1α信号通路调控糖酵解探讨阳和汤干预乳腺癌肺转移皮下瘤的机制  

Mechanism of Yanghe Decoction against subcutaneous tumor in pulmonary metastasis from breast cancer through HIF-1αsignaling pathway regulating glycolysis:based on network pharmacology and animal experiment

作　　者：刘洋靖 李晓柳 马朝群[1] 陈德轩[1] 王高元[1] 朱太阳 LIU Yang-jing;LI Xiao-liu;MA Chao-qun;CHEN De-xuan;WANG Gao-yuan;ZHU Tai-yang(Jiangsu Provincial Hospital of Chinese Medicine,Nanjing 210029,China;Nanjing Hospital of Chinese Medicine,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210022,China)

机构地区：[1]江苏省中医院,江苏南京210029 [2]南京中医药大学附属医院南京中医院,江苏南京210022

出　　处：《中国中药杂志》2023年第9期2352-2359,共8页China Journal of Chinese Materia Medica

基　　金：江苏省科技计划项目(BE2022819);江苏省中医院创新发展基金专项(MS2021019)。

摘　　要：采用网络药理学和动物实验研究阳和汤干预乳腺癌肺转移皮下瘤的作用机制,为探讨阳和汤治疗乳腺癌提供实验依据和参考。于TCMSP(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform)和SwissTargetPrediction数据库检索阳和汤组方药物的化学成分并获取靶点;GeneCards、OMIM(Online Mendelian Inheritance in Man)数据库查询疾病靶点;Excel筛选交集靶点并绘制韦恩图;构建蛋白互作网络;R语言进行基因本体论(Gene Ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析。53只SPF级雌性Bablc/6小鼠随机分为正常组、模型组以及阳和汤低、高剂量组,正常组8只,其余每组15只,阳和汤灌胃30 d,模型组、正常组灌胃同体积生理盐水。每日称量体质量、测量瘤体大小等情况,绘制小鼠体质量变化曲线、原位瘤生长曲线,结束后收集乳腺癌皮下瘤样本,HE染色观察皮下瘤,PCR及Western blot检测缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)、丙酮酸激酶M2(pyruvate kinase M2,PKM2)、乳酸脱氢酶A(lactate dehydrogenase A,LDHA)、葡萄糖转运子1(glucose transporter 1,GLUT1)的mRNA及蛋白水平。网络药理学获取阳和汤213个活性成分、185个药物-疾病交集靶点,并提出阳和汤可能通过HIF-1α信号通路调控糖酵解干预乳腺癌的假设。动物实验证实,与模型组比较,阳和汤高、低剂量组HIF-1α、PKM2、LDHA、GLUT1的mRNA及蛋白表达水平均显著降低。综上,阳和汤早期对乳腺癌肺转移皮下瘤具有一定的抑制作用,可能通过HIF-1α信号通路调控糖酵解对乳腺癌肺转移进行干预。This study aims to explore the mechanism of Yanghe Decoction(YHD)against subcutaneous tumor in pulmonary metastasis from breast cancer,which is expected to lay a basis for the treatment of breast carcinoma with YHD.The chemical components of medicinals in YHD,and the targets of the components were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and SwissTargetPrediction.The disease-related targets were searched from GeneCards and Online Mendelian Inheritance in Man(OMIM).Excel was employed to screen the common targets and plot the Venn diagram.The protein-protein interaction network was constructed.R language was used for Gene Ontology(GO)term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment.A total of 53 female SPF Bablc/6 mice were randomized into normal group(same volume of normal saline,ig),model group(same volume of normal saline,ig),and low-dose and high-dose YHD groups(YHD,ig,30 days),with 8 mice in normal group and 15 mice in each of the other groups.Body weight and tumor size was measured every day.Curves for body weight variation and growth of tumor in situ were plotted.In the end,the subcutaneous tumor sample was collected and observed based on hematoxylin and eosin(HE)staining.The mRNA and protein levels of hypoxia inducible factor-1α(HIF-1α),pyruvate kinase M2(PKM2),lactate dehydrogenase A(LDHA),and glucose transporter type 1(GLUT1)were detected by PCR and Western blot.A total of 213 active components of YHD and 185 targets against the disease were screened out.The hypothesis that YHD may regulate glycolysis through HIF-1αsignaling pathway to intervene in breast cancer was proposed.Animal experiment confirmed that the mRNA and protein levels of HIF-1α,PKM2,LDHA,and GLUT1 in the high-and low-dose YHD groups were lower than those in the model group.YHD has certain inhibitory effect on subcutaneous tumor in pulmonary metastasis from breast cancer in the early stage,which may intervene pulmonary metastasis from breast

关 键 词：乳腺癌肺转移 阳和汤 HIF-1α信号通路 糖酵解 网络药理学 

分 类 号：R285.5[医药卫生—中药学]