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.
Nanjing University of Chinese Medicine Nanjing 210029, China Affiliated Hospital of Nanjing University of Chinese Medicine Nanjing 210029, China.
Zhongguo Zhong Yao Za Zhi. 2024 Nov;49(21):5762-5770. doi: 10.19540/j.cnki.cjcmm.20240709.401.
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 concentration-dependent manner. The results indicated that in the colorectal cancer inflammatory microenvironment, the herb pair Agrimoniae Herba-Coptidis Rhizoma could inhibit angiogenesis via multiple components, targets, and pathways. The anti-angiogenesis effect might be related to the down-regulation of the expression levels of angiogenesis-related factors VEGFA, kdrl, and Flt4 in the VEGFA/VEGFR2 signaling pathway.
本研究旨在运用网络药理学方法和斑马鱼模型，探讨中药药对仙鹤草 - 黄连在抑制结直肠癌炎症微环境中血管生成的作用及机制。采用网络药理学方法获取药对在抑制结直肠癌炎症微环境血管生成中所调控的活性成分、潜在核心靶点及信号通路，并在此基础上预测其潜在机制。建立结直肠癌斑马鱼模型并模拟炎症微环境，观察不同浓度药对对斑马鱼模型节间血管（ISV）面积、数量及长度的影响。采用蛋白质免疫印迹法和实时定量聚合酶链反应分别检测血管内皮生长因子A（VEGFA）、血管内皮生长因子受体2（VEGFR2，又称kdrl、Flk1）和血管内皮生长因子受体3（VEGFR3，又称Flt4）的蛋白质和mRNA水平。共预测出仙鹤草 - 黄连18种活性成分和488个潜在靶点，药对与疾病共有108个共同靶点。根据KEGG通路富集分析结果，选取VEGFA/VEGFR2信号通路中与血管生成相关的因子VEGFA、kdrl和Flt4进行验证。斑马鱼实验表明，与空白组相比，模型组在炎症微环境中ISV的面积、数量及长度增加。与模型组相比，药对呈浓度依赖性降低ISV的面积、数量及长度。与空白组相比，模型组在炎症微环境中VEGFA、kdrl和Flt4的蛋白质和mRNA水平上调。与模型组相比，药对呈浓度依赖性下调VEGFA、kdrl和Flt4的蛋白质和mRNA水平。结果表明，在结直肠癌炎症微环境中，仙鹤草 - 黄连药对可通过多种成分、靶点和通路抑制血管生成。其抗血管生成作用可能与下调VEGFA/VEGFR2信号通路中血管生成相关因子VEGFA、kdrl和Flt4的表达水平有关。
