Liao Dan, Wang Jie, Zhang Xiaoyu, Li Rong, Yang Xiaoli
Department of Ophthalmology, Affiliated Hospital of North Sichuan Medical College, Nanchong, Sichuan, China.
Medical School of Ophthalmology and Optometry, North Sichuan Medical College, Nanchong, Sichuan, China.
Invest Ophthalmol Vis Sci. 2025 Jan 2;66(1):58. doi: 10.1167/iovs.66.1.58.
Ocular neovascularization is a major cause of blindness. Although fibroblast growth factor-2 (FGF2) has been implicated in the pathophysiology of angiogenesis, the underlying mechanisms remain incompletely understood. The purpose of this study was to investigate the role of FGF2 in retinal neovascularization and elucidate its underlying mechanisms.
The oxygen-induced retinopathy mouse model was used to study the pathogenesis of retinal neovascularization. Immunofluorescence was used to quantify the neovascularization in retina. Data-independent acquisition proteomics were performed to quantify differentially expressed proteins in human retinal microvascular endothelial cells stimulated with FGF2 and associated pathways were analyzed. We carried out qRT-PCR and Western Blot assays to detect the expression of genes at mRNA and protein levels. The angiogenesis abilities of human retinal microvascular endothelial cells were measured by transwell, EdU and tube formation assays.
FGF2 was significantly upregulated in retinal tissues of the oxygen-induced retinopathy mouse model and it markedly enhanced tube formation, migration, and proliferation abilities of human retinal microvascular endothelial cells in vitro. The proteomic analysis identified 287 differentially expressed proteins in endothelial cells in response to FGF2 stimulation, characterized by a notable upregulation of the glycolysis pathway, among which we confirmed that the enolase 2 (ENO2) levels were elevated after FGF2 stimulation, and its knockdown resulted in diminished glycolytic activity and impaired angiogenic processes. Furthermore, the use of the ENO2 inhibitor AP-Ⅲ-a4 alleviated angiogenesis in vivo and in vitro.
Our findings underscore the pivotal role of ENO2-mediated glycolysis in FGF2-induced angiogenesis, suggesting that ENO2 may serve as a promising therapeutic target for managing pathological neovascularization.
眼部新生血管形成是失明的主要原因。尽管成纤维细胞生长因子-2（FGF2）与血管生成的病理生理学有关，但其潜在机制仍不完全清楚。本研究的目的是探讨FGF2在视网膜新生血管形成中的作用，并阐明其潜在机制。
采用氧诱导视网膜病变小鼠模型研究视网膜新生血管形成的发病机制。免疫荧光法用于量化视网膜中的新生血管形成。进行数据非依赖采集蛋白质组学以量化用FGF2刺激的人视网膜微血管内皮细胞中差异表达的蛋白质，并分析相关途径。我们进行了qRT-PCR和蛋白质免疫印迹分析以检测基因在mRNA和蛋白质水平的表达。通过Transwell、EdU和管形成试验测量人视网膜微血管内皮细胞的血管生成能力。
在氧诱导视网膜病变小鼠模型的视网膜组织中，FGF2显著上调，并且它在体外显著增强了人视网膜微血管内皮细胞的管形成、迁移和增殖能力。蛋白质组学分析鉴定出内皮细胞中287种响应FGF2刺激的差异表达蛋白质，其特征在于糖酵解途径显著上调，其中我们证实FGF2刺激后烯醇化酶2（ENO2）水平升高，并且其敲低导致糖酵解活性降低和血管生成过程受损。此外，使用ENO2抑制剂AP-Ⅲ-a4可减轻体内和体外的血管生成。
我们的研究结果强调了ENO2介导的糖酵解在FGF2诱导的血管生成中的关键作用，表明ENO2可能作为治疗病理性新生血管形成的有前景的治疗靶点。
