Liang Yan, Su Tingting, Zhu Shijiao, Sun Ruolan, Qin Jiahui, Yue Zengyaran, Wang Xu, Liang Zhongqing, Tan Xiying, Bian Yong, Zhao Fan, Tang Decai, Yin Gang
School of Chinese Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, China.
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023, China.
Phytomedicine. 2025 May;140:156562. doi: 10.1016/j.phymed.2025.156562. Epub 2025 Feb 24.
Abnormal tumor blood vessels can significantly promote the malignant progression of tumors, prompting researchers to focus on drugs that normalize these vessels for clinical treatment. The combination of the Qi-tonifying drug Astragali Radix and the blood-activating drug Curcumae Rhizoma, referred to as AC, exhibited significant anti-tumor metastasis effects. However, the association between the anti-tumor metastasis effect of AC and its potential role in regulating tumor vascular remodeling warrants further exploration.
This study aimed to elucidate the mechanism through which AC induces tumor blood vessel normalization in colon cancer (CC).
The potential active components of AC were identified through UPLC-MS/MS. An orthotopic transplantation model of CC was established in BALB/c mice using the CT26-Lucifer cell line, and the effects of AC were evaluated using IVIS imaging, hematoxylin and eosin (H&E) staining, and immunohistochemistry. Network pharmacology and molecular biology analyses were employed to identify the potential direct targets of AC. Subsequently, RT-PCR and Western blotting techniques were utilized to validate the findings obtained from network pharmacology. Furthermore, ELISA and other methodologies were used to investigate glycolysis-related indicators, along with immunofluorescence technology to demonstrate changes in vascular leakage and perfusion characteristics associated with blood vessel normalization.
We identified HIF-1α as a potential direct target of AC. This interaction influences the glycolytic processes in both tumor cells and tumor-associated endothelial cells (TECs) by directly binding to HIF-1α and modulating its nuclear translocation, thereby determining the integrity of TEC junctions. Mechanistically, AC directly regulates the key enzyme PFKFB3 in glycolysis by modulating HIF-1α expression and inhibiting its nuclear translocation. This action reduces tumor glycolytic flux, decreases the internalization of VE-cad, and influences the expression of downstream matrix metalloproteinases (MMPs), thereby strengthening the adherens and tight junctions between TECs and restoring vascular integrity.
This study presents novel findings that AC can regulate glycolysis through the inhibition of HIF-1α nuclear translocation, thereby promoting the normalization of tumor blood vessels and effectively inhibiting tumor metastasis. These results suggested that AC may serve as an effective therapeutic agent for normalizing tumor blood vessels.
异常的肿瘤血管可显著促进肿瘤的恶性进展，促使研究人员关注使这些血管正常化的药物用于临床治疗。补气药黄芪与活血药莪术的组合，称为AC，表现出显著的抗肿瘤转移作用。然而，AC的抗肿瘤转移作用与其在调节肿瘤血管重塑中的潜在作用之间的关联值得进一步探索。
本研究旨在阐明AC诱导结肠癌（CC）肿瘤血管正常化的机制。
通过超高效液相色谱-串联质谱（UPLC-MS/MS）鉴定AC的潜在活性成分。使用CT26-Lucifer细胞系在BALB/c小鼠中建立CC原位移植模型，并使用IVIS成像、苏木精和伊红（H&E）染色以及免疫组织化学评估AC的作用。采用网络药理学和分子生物学分析来鉴定AC的潜在直接靶点。随后，利用逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹技术验证从网络药理学获得的结果。此外，采用酶联免疫吸附测定（ELISA）等方法研究糖酵解相关指标，并利用免疫荧光技术证明与血管正常化相关的血管渗漏和灌注特征的变化。
我们确定缺氧诱导因子-1α（HIF-1α）为AC的潜在直接靶点。这种相互作用通过直接结合HIF-1α并调节其核转位来影响肿瘤细胞和肿瘤相关内皮细胞（TECs）中的糖酵解过程，从而决定TEC连接的完整性。机制上，AC通过调节HIF-1α表达并抑制其核转位来直接调节糖酵解中的关键酶6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3（PFKFB3）。这一作用降低肿瘤糖酵解通量，减少血管内皮钙黏蛋白（VE-cad）的内化，并影响下游基质金属蛋白酶（MMPs）的表达，从而加强TECs之间的黏附连接和紧密连接并恢复血管完整性。
本研究提出了新的发现，即AC可通过抑制HIF-1α核转位来调节糖酵解，从而促进肿瘤血管正常化并有效抑制肿瘤转移。这些结果表明AC可能是一种使肿瘤血管正常化的有效治疗剂。
