Centro de Investigaciones en Bioquímica Clínica e Inmunología (CIBICI-CONICET), Departamento de Bioquímica Clínica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Córdoba, Argentina.
The Wilmer Eye Institute, Johns Hopkins University, Baltimore, MD, USA.
Mol Neurobiol. 2018 Feb;55(2):1123-1135. doi: 10.1007/s12035-017-0386-9. Epub 2017 Jan 17.
In ischemic proliferative diseases such as retinopathies, persistent hypoxia leads to the release of numerous neovascular factors that participate in the formation of abnormal vessels and eventually cause blindness. The upregulation and activation of metalloproteinases (MMP-2 and MMP-9) represent a final common pathway in this process. Although many regulators of the neovascular process have been identified, the complete role of the insulin-like growth factor 1 (IGF-1) and its receptor (IGF-1R) appears to be significantly more complex. In this study, we used an oxygen-induced retinopathy (OIR) mouse model as well as an in vitro model of hypoxia to study the role of MMP-2 derived from Müller glial cells (MGCs) and its relation with the IGF-1/IGF-1R system. We demonstrated that MMP-2 protein expression increased in P17 OIR mice, which coincided with the active phase of the neovascular process. Also, glutamine synthetase (GS)-positive cells were also positive for MMP-2, whereas IGF-1R was expressed by GFAP-positive cells, indicating that both proteins were expressed in MGCs. In addition, in the OIR model a single intravitreal injection of the IGF-1R blocking antibody (αIR3) administered at P12 effectively prevented pathologic neovascularization, accelerated physiological revascularization, and improved retinal functionality at P17. Finally, in MGC supernatants, the blocking antibody abolished the IGF-1 effect on active MMP-2 under normoxic and hypoxic conditions without affecting the extracellular levels of pro-MMP-2. These results demonstrate, for the first time, that the IGF-1/IGF-1R system regulates active MMP-2 levels in MGCs, thus contributing to MEC remodeling during the retinal neovascular process.
在缺血性增殖性疾病(如视网膜病变)中,持续缺氧会导致大量新生血管因子的释放,这些因子参与异常血管的形成,最终导致失明。金属蛋白酶(MMP-2 和 MMP-9)的上调和激活是这个过程的最后共同途径。尽管已经确定了许多新生血管过程的调节剂,但胰岛素样生长因子 1(IGF-1)及其受体(IGF-1R)的完整作用似乎要复杂得多。在这项研究中,我们使用氧诱导的视网膜病变(OIR)小鼠模型以及缺氧体外模型来研究来自 Müller 胶质细胞(MGC)的 MMP-2 的作用及其与 IGF-1/IGF-1R 系统的关系。我们证明,在 P17 OIR 小鼠中 MMP-2 蛋白表达增加,这与新生血管过程的活跃阶段相吻合。此外,谷氨酰胺合成酶(GS)阳性细胞也呈 MMP-2 阳性,而 IGF-1R 则由 GFAP 阳性细胞表达,表明这两种蛋白均在 MGC 中表达。此外,在 OIR 模型中,在 P12 时单次玻璃体内注射 IGF-1R 阻断抗体(αIR3)可有效预防病理性新生血管形成,加速生理性血管再通,并在 P17 时改善视网膜功能。最后,在 MGC 上清液中,阻断抗体在正常氧和缺氧条件下可消除 IGF-1 对活性 MMP-2 的作用,而不影响细胞外 pro-MMP-2 的水平。这些结果首次证明,IGF-1/IGF-1R 系统调节 MGC 中活性 MMP-2 的水平,从而有助于视网膜新生血管过程中 MEC 的重塑。