Department of Ophthalmology, Henry Ford Health System, Detroit, Michigan, United States Department of Ophthalmology, Shaanxi Maternity and Child Healthcare Hospital, Xi'an, Shaanxi, People's Republic of China.
Department of Ophthalmology, Henry Ford Health System, Detroit, Michigan, United States.
Invest Ophthalmol Vis Sci. 2014 Jun 26;55(7):4461-9. doi: 10.1167/iovs.14-14451.
The redox function of APE1/Ref-1 is a key regulator in pathological angiogenesis, such as retinal neovascularization and tumor growth. In this study, we examined whether inhibition of APE1/Ref-1 redox function by a small molecule inhibitor E3330 suppresses experimental choroidal neovascularization (CNV) in vitro and in vivo.
Primate choroid endothelial cells (CECs) received treatment of 0 to 100 μM E3330 alone or cotreatment of E3330 and 500 μg/mL anti-VEGF antibody bevacizumab. Choroid endothelial cell angiogenic function was examined by cell proliferation, migration, and tube formation assays. The effects of E3330 on NF-κB and STAT3 signaling pathways were determined by reporter gene assay, Western blot, and ELISA. Laser-induced CNV mouse model was used to test the effects of E3330 in vivo. Potential toxicity of E3330 was evaluated by TUNEL assay.
The E3330 of 25 to 100 μM dose-dependently suppressed CEC proliferation, migration, and tube formation, in the absence of noticeable cell toxicity. Lower doses of E3330 (10-20 μM) reduced the transcriptional activity of NF-κB and STAT3 without affecting protein phosphorylation of both molecules. At the same time, E3330 downregulated MCP-1 production in CECs. The antiangiogenic effect of E3330 was comparable and additive to bevacizumab. The E3330 effectively attenuated the progression of laser-induced CNV in mice after a single intravitreal injection.
The APE1/Ref-1 redox function regulates multiple transcription factors and inflammatory molecules, and is essential for CEC angiogenesis. Specific inhibition of APE1/Ref-1 redox function with E3330 may represent a promising novel treatment for wet AMD.
APE1/Ref-1 的氧化还原功能是病理性血管生成的关键调节因子，如视网膜新生血管和肿瘤生长。在这项研究中，我们研究了小分子抑制剂 E3330 抑制 APE1/Ref-1 氧化还原功能是否能抑制体外和体内实验性脉络膜新生血管（CNV）。
灵长类脉络膜内皮细胞（CEC）接受 0 至 100μM E3330 单独或 E3330 与 500μg/ml 抗血管内皮生长因子抗体贝伐单抗联合处理。通过细胞增殖、迁移和管形成试验检测脉络膜内皮细胞的血管生成功能。通过报告基因检测、Western blot 和 ELISA 测定 E3330 对 NF-κB 和 STAT3 信号通路的影响。采用激光诱导的 CNV 小鼠模型体内检测 E3330 的作用。通过 TUNEL 试验评价 E3330 的潜在毒性。
25 至 100μM 剂量的 E3330 呈剂量依赖性抑制 CEC 增殖、迁移和管形成，而无明显细胞毒性。较低剂量的 E3330（10-20μM）降低 NF-κB 和 STAT3 的转录活性，而不影响两种分子的蛋白磷酸化。同时，E3330 下调 CEC 中 MCP-1 的产生。E3330 的抗血管生成作用与贝伐单抗相当且具有相加作用。单次玻璃体内注射 E3330 可有效抑制激光诱导的 CNV 在小鼠中的进展。
APE1/Ref-1 的氧化还原功能调节多种转录因子和炎症分子，对 CEC 血管生成至关重要。APE1/Ref-1 氧化还原功能的特异性抑制可能为湿性 AMD 提供一种有前景的新治疗方法。
