基于网络药理学和实验验证研究仙鹤草-黄连药对抗结直肠癌炎性微环境中血管生成活性  

Agrimoniae Herba-Coptidis Rhizoma inhibits angiogenesis in colorectal cancer inflammatory microenvironment based on network pharmacology and experiment validation

作　　者：沈鑫玲 彭海燕[1,2] 符黄杰 何亚萍 李智育 侯敏艳 张淑娟 熊晗 SHEN Xin-ling;PENG Hai-yan;FU Huang-jie;HE Ya-ping;LI Zhi-yu;HOU Min-yan;ZHANG Shu-juan;XIONG Han(Nanjing University of Chinese Medicine,Nanjing 210029,China;Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]南京中医药大学,江苏南京210029 [2]南京中医药大学附属医院,江苏南京210029 [3]浙江中医药大学,浙江杭州310053

出　　处：《中国中药杂志》2024年第21期5762-5770,共9页China Journal of Chinese Materia Medica

基　　金：江苏省自然科学基金项目(BK20201400);全国中医药研究生教育课题(YJS-YB-2023-49);江苏高校优势学科建设工程项目(苏政办发[2018]87号);南京中医药大学名老中医药专家学术传承首批建设项目(教字[2022]44号)。

摘　　要：基于网络药理学和斑马鱼体内实验,探讨结直肠癌炎性微环境中仙鹤草-黄连药对对血管生成的影响和潜在机制。通过网络药理学获得结直肠癌炎性微环境中仙鹤草-黄连药对抗血管生成的有效活性成分、潜在核心靶点和信号通路,预测结直肠癌炎性微环境中仙鹤草-黄连药对抑制血管生成的作用机制;建立斑马鱼结直肠癌模型,并进行炎性微环境造模,观察不同浓度的仙鹤草-黄连药对对斑马鱼体节间血管(ISVs)面积、数量、长度的影响,蛋白免疫印迹法检测斑马鱼血管内皮生长因子A(VEGFA)、血管生长因子受体2(VEGFR2,又名kdrl、Flk1)、血管生长因子受体3(VEGFR3,又名Flt4)的蛋白表达水平,实时荧光定量聚合酶链式反应检测斑马鱼VEGFA、kdrl、Flt4 mRNA的表达水平。网络药理学筛选出仙鹤草-黄连药对中18个活性成分和488个潜在靶点,药物和疾病交集靶点108个。结合KEGG通路富集分析,选择VEGFA/VEGFR2信号通路中血管生成相关因子VEGFA、kdrl、Flt4进行验证。斑马鱼实验表明,与空白组比较,模型组斑马鱼ISVs面积、数量、长度均增加;与模型组比较,仙鹤草-黄连药对各剂量组斑马鱼ISVs面积、数量、长度均减少,且具有浓度依赖性。与空白组比较,模型组斑马鱼VEGFA、kdrl、Flt4蛋白和mRNA表达水平均上调;与模型组比较,仙鹤草-黄连药对各剂量组斑马鱼VEGFA、kdrl、Flt4蛋白和mRNA表达水平均下调,且具有浓度依赖性。结果表明,在结直肠癌炎性微环境中,仙鹤草-黄连药对能够通过多成分、多靶点、多通路抑制肿瘤血管生成,且结直肠癌炎性微环境下其抗血管生成作用可能与下调VEGFA/VEGFR2通路中血管生成相关因子VEGFA、kdrl、Flt4表达水平有关。This study aims to investigate the effect and mechanism of the herb pair Agrimoniae Herba-Coptidis Rhizoma in inhibiting angiogenesis in the colorectal cancer inflammatory microenvironment by using the method of network pharmacology and the zebrafish model.The method of network pharmacology was employed to obtain the active components,potential core targets,and signaling pathways regulated by the herb pair in inhibiting angiogenesis in the inflammatory microenvironment of colorectal cancer,on the basis of which the underlying mechanism was predicted.The zebrafish model of colorectal cancer was established,and the inflammatory microenvironment was modeled.The effects of different concentrations of the herb pair on the area,number,and length of intersegmental vessels(ISVs)of the zebrafish model were observed.Western blot and real-time quantitative PCR were employed to measure the protein and mRNA levels,respectively,of vascular endothelial growth factor A(VEGFA),vascular epidermal growth factor receptor 2(VEGFR2,also known as kdrl,Flk1),and vascular epidermal growth factor receptor 3(VEGFR3,also known as Flt4).A total of 18 active components and 488 potential targets of Agrimoniae Herba-Coptidis Rhizoma were predicted,and 108 common targets were shared by the herb pair and the disease.According to the results of KEGG pathway enrichment analysis,the angiogenesis-related factors VEGFA,kdrl,and Flt4 in the VEGFA/VEGFR2 signaling pathway were selected for verification.The zebrafish experiment showed that compared with the blank group,the model group showed increased area,number,and length of ISVs in the inflammatory microenvironment.Compared with the model group,the herb pair decreased the area,number,and length of ISVs in a concentration-dependent manner.Compared with the blank group,the model group showed up-regulated protein and mRNA levels of VEGFA,kdrl,and Flt4 in the inflammatory microenvironment.Compared with the model group,the herb pair down-regulated the protein and mRNA levels of VEGFA,kdrl,and Flt4 in a con

关 键 词：仙鹤草-黄连 结直肠癌 斑马鱼 网络药理学 炎性微环境 血管生成 血管生成相关因子 作用机制 

分 类 号：R285.5[医药卫生—中药学]