Fan Yuan-Yao, Liu Chi-Hsien, Wu An-Lun, Chen Hung-Chi, Hsueh Yi-Jen, Chen Kuan-Jen, Lai Chi-Chun, Huang Chung-Ying, Wu Wei-Chi
Department of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan, Taiwan; Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
Graduate Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan, Taiwan.
Eur J Pharmacol. 2021 Jun 5;900:174035. doi: 10.1016/j.ejphar.2021.174035. Epub 2021 Mar 13.
Vascular endothelial growth factor (VEGF) is the principal growth factor responsible for the retinal neovascularization in the pathogenesis of retinopathy of prematurity (ROP). Current therapies for ROP include laser ablation and intravitreal anti-VEGF injection. However, these treatments either destroy the peripheral retina or associate with problems of persistent peripheral avascular retina or later recurrence of ROP. In the present study we investigated a new therapeutic approach by exploring the potential role of a specific microRNA, miR-126, in regulating VEGFA expression and retinal neovascularization in a rat oxygen-induced retinopathy (OIR) model. We demonstrated that miR-126 mimic and plasmid effectively suppresses VEGFA mRNA expression in both human and rat retinal pigment epithelium cell lines, quantified with qRT-PCR. Animal experiments on rat OIR model revealed that intravitreal injection of miR-126 plasmid efficiently downregulated VEGFA expression in the intraocular fluid and retinal tissues measured by ELISA, and significantly suppressed retinal neovascularization, which was confirmed by calculating sizes of neovascularization areas on fluorescence microscopic images of flat mounted retina stained with Alexa Fluor 594-conjugated isolectin B4 to visualize blood vessels. Together, these results showed that intravitreal injection of miR-126 plasmid could inhibit retinal neovascularization by down-regulating VEGFA expression, suggesting a potential therapeutic effect for ROP.
血管内皮生长因子（VEGF）是早产儿视网膜病变（ROP）发病机制中导致视网膜新生血管形成的主要生长因子。目前针对ROP的治疗方法包括激光消融和玻璃体内注射抗VEGF药物。然而，这些治疗方法要么会破坏周边视网膜，要么会出现周边视网膜持续无血管化或ROP后期复发的问题。在本研究中，我们通过探索一种特定的微小RNA，即miR-126，在大鼠氧诱导性视网膜病变（OIR）模型中调节VEGFA表达和视网膜新生血管形成的潜在作用，研究了一种新的治疗方法。我们证明，用qRT-PCR定量分析显示，miR-126模拟物和质粒可有效抑制人及大鼠视网膜色素上皮细胞系中VEGFA mRNA的表达。对大鼠OIR模型进行的动物实验表明，通过ELISA检测，玻璃体内注射miR-126质粒可有效下调眼内液和视网膜组织中VEGFA的表达，并显著抑制视网膜新生血管形成，这通过计算用Alexa Fluor 594偶联的异凝集素B4染色的扁平视网膜荧光显微镜图像上的新生血管区域大小得以证实，该方法用于可视化血管。总之，这些结果表明，玻璃体内注射miR-126质粒可通过下调VEGFA表达来抑制视网膜新生血管形成，提示其对ROP具有潜在治疗作用。
