基于VEGF/ANG通路探究固本抑瘤Ⅱ号对三阴性乳腺癌术后自发肺转移的作用机制

左曦; 王笑民; 南凌杉; 殷晓辉; 陈栋; 张甘霖

doi:10.16025/j.1674-1307.2026.04.005

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基于VEGF/ANG通路探究固本抑瘤Ⅱ号对三阴性乳腺癌术后自发肺转移的作用机制

实验研究 | 更新时间：2026-06-10

- 基于VEGF/ANG通路探究固本抑瘤Ⅱ号对三阴性乳腺癌术后自发肺转移的作用机制
- Mechanisms of Guben Yiliu No. 2 in inhibiting postoperative lung metastasis of triple-negative breast cancer mice via the VEGF/ANG pathway
- 北京中医药 2026年45卷第4期页码：452-460
- 作者机构：
  
  1.北京中医药大学，北京 100029
  2.首都医科大学附属北京中医医院肿瘤科，北京 100010
- 作者简介：
  
  左曦，女，29岁，博士研究生。研究方向：中西医结合防治恶性肿瘤。
  张甘霖，E-mail：kalinezhang@163.com
- 基金信息：
  
  国家自然科学基金项目(82274599)
- DOI：10.16025/j.1674-1307.2026.04.005
  中图分类号：
- 收稿：2025-10-30，
  
  纸质出版：2026-04-25
- 稿件说明：
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左曦，王笑民，南凌杉，等.基于VEGF/ANG通路探究固本抑瘤Ⅱ号对三阴性乳腺癌术后自发肺转移的作用机制［J］.北京中医药，2026，45（4）：452-460.

ZUO Xi,WANG Xiaomin,NAN Lingshan,et al.Mechanisms of Guben Yiliu No. 2 in inhibiting postoperative lung metastasis of triple-negative breast cancer mice via the VEGF/ANG pathway[J]. Beijing Journal of Traditional Chinese Medicine,2026,45(04):452-460.
左曦，王笑民，南凌杉，等.基于VEGF/ANG通路探究固本抑瘤Ⅱ号对三阴性乳腺癌术后自发肺转移的作用机制［J］.北京中医药，2026，45（4）：452-460. DOI： 10.16025/j.1674-1307.2026.04.005.

ZUO Xi,WANG Xiaomin,NAN Lingshan,et al.Mechanisms of Guben Yiliu No. 2 in inhibiting postoperative lung metastasis of triple-negative breast cancer mice via the VEGF/ANG pathway[J]. Beijing Journal of Traditional Chinese Medicine,2026,45(04):452-460. DOI： 10.16025/j.1674-1307.2026.04.005.

摘要

目的

探究固本抑瘤Ⅱ号通过血管内皮生长因子/血管生成素（VEGF/ANG）通路减少肿瘤血管生成，进而抑制三阴性乳腺癌（TNBC）小鼠术后肺转移的作用和机制。

方法

将25只雌性Balb/c小鼠，随机分为空白组、模型组、术后中药组、全程中药组、表柔比星组，每组5只。采用4T1-luciferase细胞第4乳垫原位接种+术后切除原位瘤法构建TNBC术后自发肺转移模型，术后中药组于术后第3天，全程中药组于接种后第3天开始用固本抑瘤Ⅱ号［60 mL/（kg·d）］灌胃，表柔比星组腹腔注射表柔比星（0.003 g/kg，1周2次），空白组与模型组给予等体积生理盐水。通过小动物活体可见光成像、micro-CT评估肺转移情况。免疫组织化学血小板内皮细胞黏附分子-1/过碘酸-雪夫染色（CD31/PAS）检测血管生成拟态；蛋白免疫印迹检测肺组织血管内皮生长因子A（VEGFA）、血管生成素-1（ANG1）、血管生成素-2（ANG2）、酪氨酸激酶受体2（Tie2）等蛋白表达；酶联免疫吸附试验（ELISA）检测血清VEGF水平；免疫荧光检测肺组织紧密连接蛋白-1（claudin-1）表达。通过体外肺微血管内皮细胞（PMVEC）与4T1细胞共培养，检测细胞成管能力及血管通透性。

结果

与模型组比较，全程中药组肺转移灶荧光强度、肺质量及肺指数均显著降低（

0.05），肺组织MVD、VM密度显著下降（

0.05），claudin-1荧光强度显著增强（

0.05）；血清VEGF及肺组织VEGFA、ANG2蛋白表达显著下调（

0.05），ANG1及Tie2表达呈上升趋势。体外实验中，固本抑瘤Ⅱ号显著抑制PMVEC成管结构数、成管长度及节点数（

0.05），降低FITC-葡聚糖渗漏量（

0.05），而表柔比星反而增加其渗透量。

结论

固本抑瘤Ⅱ号可有效抑制TNBC小鼠术后肺转移，其机制可能是通过调控VEGF/ANG通路，抑制肿瘤血管生成、降低肺微血管通透性，从而阻止转移前生态位形成。

Abstract

Objective To investigate the effects and mechanisms of

Guben Yiliu

No. 2 in inhibiting postoperative lung metastasis of triple-negative breast cancer （TNBC） in mice by reducing tumor angiogenesis through the vascular endothelial growth factor/angiopoietin （VEGF/ANG） pathway. Methods Twenty-five female BALB/c mice were randomly divided into a blank group， model group， postoperative traditional Chinese medicine （TCM） group， whole-course TCM group， and epirubicin group， with 5 mice in each group. A spontaneous postoperative lung metastasis model of TNBC was established by orthotopic inoculation of 4T1-luciferase cells into the fourth mammary fat pad followed by surgical resection of the primary tumor. The postoperative TCM group received

Guben Yiliu

No. 2 （60 mL/kg•d） by gavage starting on postoperative day 3， while the whole-course TCM group received the same treatment beginning on day 3 after tumor inoculation. The epirubicin group received intraperitoneal injections of epirubicin （0.003 g/kg， twice weekly）. The blank and model groups received equal volumes of normal saline. Lung metastasis was evaluated by small-animal in vivo bioluminescence imaging and micro-CT. Vasculogenic mimicry （VM） was detected by immunohistochemical staining for platelet endothelial cell adhesion molecule-1/periodic acid-Schiff （CD31/PAS）. Protein expression levels of vascular

endothelial growth factor A （VEGFA）， angiopoietin-1 （ANG1）， angiopoietin-2 （ANG2）， and tyrosine kinase receptor 2 （Tie2） in lung tissue were detected by Western blot. Serum VEGF levels were measured by enzyme-linked immunosorbent assay （ELISA）. Expression of the tight junction protein-1 （claudin-1） in lung tissue was detected by immunofluorescence. In vitro， pulmonary microvascular endothelial cells （PMVECs） were co-cultured with 4T1 cells to evaluate tube formation ability and vascular permeability. Results Compared with the model group， the whole-course TCM group showed significantly reduced fluorescence intensity of lung metastatic lesions， lung weight， and lung index （

0.05）， significantly decreased microvessel density （MVD） and VM density in lung tissue （

0.05）， and significantly enhanced claudin-1 fluorescence intensity （

0.05）. Serum VEGF levels and protein expression levels of VEGFA and ANG2 in lung tissue were significantly downregulated （

0.05）， whereas ANG1 and Tie2 expression levels showed an upward trend. In vitro experiments demonstrated that

Guben Yiliu

No. 2 significantly inhibited the number of PMVEC tube structures， tube length， and node number （

0.05）， and reduced FITC-dextran leakage （

0.05）， whereas epirubicin increased vascular permeability. Conclusion

Guben Yiliu

No. 2 effectively inhibits postoperative lung metastasis in TNBC mice. The underlying mechanism may involve regulation of the VEGF/ANG pathway， inhibition of tumor angiogenesis， and reduction of pulmonary microvascular permeability， thereby preventing the formation of the premetastatic niche.

关键词

Keywords

references

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