Li Zuohong, Chen Jianping, Huang Zhaohao, Huang Weifeng, Wang Kerui, Liang Xuanwei, Su Wenru
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
Exp Eye Res. 2025 Jan;250:110165. doi: 10.1016/j.exer.2024.110165. Epub 2024 Nov 19.
Corneal neovascularization (CNV) is a dynamically regulated process that arises due to a disruption in the equilibrium between pro-angiogenic and anti-angiogenic factors. Various cytokines are released by vascular endothelial cells and macrophages in damaged cornea, ultimately inducing CNV. The cAMP-response element-binding protein (CREB), a nuclear transcription factor, potentially impacts tumor angiogenesis by modulating the secretion of angiogenic proteins. This study aimed to assess the impact of 666-15, a potent inhibitor of CREB, on angiogenesis using human microvascular retinal endothelial cells (HMRECs), RAW 264.7 macrophage cell line and alkali-induce CNV mouse model. In vivo, the topical application of 666-15 (0.05 mg/mL) to the alkali-burn corneas led to 45% reduction in CNV. Additionally, in vitro treatment with 666-15 is effective in suppressing the migration, proliferation, and tube formation by HMRECs. Furthermore, treatment with 666-15 resulted in a down-regulation of pro-angiogenic cytokines expression, including VEGF-A, TGF-β1, b-FGF, and MMP-2 but simultaneously increasing anti-angiogenic cytokines expression, such as ADAMTS-1, Thrombospondin-1 (Tsp-1) and Tsp-2, both in alkali-burn corneas and HMRECs. And 666-15 inhibited the recruitment and the cytokines expression (VEGF-A, MMP-2, IL-1β, TNF-α, MCP-1 and MIP-1) of macrophage. Our findings revealed that 666-15 may suppress the function of endothelial cells and angiogenesis by restoring the homeostasis of pro-angiogenic stimuli, suggesting its potential as a therapeutic agent in the treatment of CNV and other angiogenesis-driven diseases.
角膜新生血管形成（CNV）是一个动态调节的过程，它是由于促血管生成因子和抗血管生成因子之间的平衡被破坏而产生的。受损角膜中的血管内皮细胞和巨噬细胞会释放各种细胞因子，最终诱导CNV。环磷酸腺苷反应元件结合蛋白（CREB）是一种核转录因子，它可能通过调节血管生成蛋白的分泌来影响肿瘤血管生成。本研究旨在使用人微血管视网膜内皮细胞（HMREC）、RAW 264.7巨噬细胞系和碱诱导的CNV小鼠模型，评估CREB的强效抑制剂666-15对血管生成的影响。在体内，将666-15（0.05 mg/mL）局部应用于碱烧伤角膜可使CNV减少45%。此外，用666-15进行体外处理可有效抑制HMREC的迁移、增殖和管形成。此外，在碱烧伤角膜和HMREC中，用666-15处理均导致促血管生成细胞因子表达下调，包括血管内皮生长因子-A（VEGF-A）、转化生长因子-β1（TGF-β1）、碱性成纤维细胞生长因子（b-FGF）和基质金属蛋白酶-2（MMP-2），但同时增加了抗血管生成细胞因子的表达，如含血小板反应蛋白基序的解聚蛋白样金属蛋白酶-1（ADAMTS-1）、血小板反应蛋白-1（Tsp-1）和血小板反应蛋白-2（Tsp-2）。并且666-15抑制了巨噬细胞的募集和细胞因子表达（VEGF-A、MMP-2、白细胞介素-1β、肿瘤坏死因子-α、单核细胞趋化蛋白-1和巨噬细胞炎性蛋白-1）。我们的研究结果表明，666-15可能通过恢复促血管生成刺激的稳态来抑制内皮细胞功能和血管生成，提示其在治疗CNV和其他血管生成驱动性疾病方面具有作为治疗剂的潜力。
