Wang Lei, Lee Amy Yi Wei, Wigg Jonathan P, Peshavariya Hitesh, Liu Ping, Zhang Hong
Eye Hospital, First Affiliated Hospital, Harbin Medical University, Harbin 150001, China.
Department of Pharmacology and Therapeutics, Drug Delivery Unit, Centre for Eye Research Australia, University of Melbourne, East Melbourne VIC 3000, Australia.
Int J Mol Sci. 2016 Jun 7;17(6):895. doi: 10.3390/ijms17060895.
miR-126 has recently been implicated in modulating angiogenic factors in vascular development. Understandings its biological significance might enable development of therapeutic interventions for diseases like age-related macular degeneration (AMD). We aimed to determine the role of miR-126 in AMD using a laser-induced choroidal neovascularization (CNV) mouse model. CNV was induced by laser photocoagulation in C57BL/6 mice. The CNV mice were transfected with scrambled miR or miR-126 mimic. The expression of miR-126, vascular endothelial growth factor-A (VEGF-A), Kinase insert domain receptor (KDR) and Sprouty-related EVH1 domain-containing protein 1 (SPRED-1) in ocular tissues were analyzed by qPCR and Western blot. The overexpression effects of miR-126 were also proven on human microvascular endothelial cells (HMECs). miR-126 showed a significant decrease in CNV mice (p < 0.05). Both mRNA and protein levels of VEGF-A, KDR and SPRED-1 were upregulated with CNV; these changes were ameliorated by restoration of miR-126 (p < 0.05). CNV was reduced after miR-126 transfection. Transfection of miR-126 reduced the HMECs 2D-capillary-like tube formation (p < 0.01) and migration (p < 0.01). miR-126 has been shown to be a negative modulator of angiogenesis in the eye. All together these results high lights the therapeutic potential of miR-126 suggests that it may contribute as a putative therapeutic target for AMD in humans.
miR-126最近被认为在调节血管发育中的血管生成因子方面发挥作用。了解其生物学意义可能有助于开发针对年龄相关性黄斑变性（AMD）等疾病的治疗干预措施。我们旨在使用激光诱导脉络膜新生血管（CNV）小鼠模型确定miR-126在AMD中的作用。通过激光光凝在C57BL/6小鼠中诱导CNV。将CNV小鼠用乱序miR或miR-126模拟物转染。通过qPCR和蛋白质印迹分析眼组织中miR-126、血管内皮生长因子-A（VEGF-A）、激酶插入结构域受体（KDR）和含Sprouty相关EVH1结构域蛋白1（SPRED-1）的表达。miR-126在人微血管内皮细胞（HMECs）上的过表达作用也得到了证实。miR-126在CNV小鼠中显著降低（p<0.05）。随着CNV的发生，VEGF-A、KDR和SPRED-1的mRNA和蛋白质水平均上调；miR-126的恢复改善了这些变化（p<0.05）。miR-126转染后CNV减少。miR-126转染减少了HMECs的二维毛细血管样管形成（p<0.01）和迁移（p<0.01）。miR-126已被证明是眼中血管生成的负调节因子。所有这些结果都突出了miR-126的治疗潜力，表明它可能作为人类AMD的潜在治疗靶点发挥作用。
