Molecular Mechanisms of SOD3 and Oxidative Stress in Diabetic Vitreoretinopathy

描述（由申请人提供）：糖尿病性玻璃体视网膜病变（DVR）是世界范围内日益增长的致盲原因。玻璃体内眼内活性氧（ROS）的升高与DVR的发病机制有关，但其分子机制尚不清楚。我们的研究表明，细胞外超氧化物歧化酶（SOD3）是人体玻璃体中一种重要的抗氧化防御酶，可以保护视网膜、睫状体和晶状体免受氧化损伤。我们建议的总体目标是研究SOD3在正常和糖尿病玻璃体中的分子机制和功能。我们的中心假设是SOD3在玻璃体亚结构上的差异定位，这些相互作用或SOD3酶活性的失调可能在DVR的病理生理中至关重要。项目目标包括：确定SOD3蛋白/蛋白多糖在人玻璃体中的相互作用机制，确定SOD3糖化在DVR中的功能作用，研究SOD3氧化应激途径在小鼠和人玻璃体中的作用。将使用几种不同的技术。首先，蛋白质相互作用将通过免疫共沉淀，免疫印迹，

DESCRIPTION (provided by applicant): Diabetic vitreoretinopathy (DVR) is a growing cause of blindness across the world. Elevation of intraocular reactive oxygen species (ROS) in the vitreous is implicated in the pathogenesis of DVR, but the molecular mechanisms are not known. Our studies suggest extracellular superoxide dismutase (SOD3) is an important antioxidant defense enzyme in the human vitreous and may protect the retina, ciliary body, and lens from oxidative damage. The overall objective of our proposal is to investigate the molecular mechanisms and function of SOD3 in the normal and diabetic vitreous. Our central hypothesis is that SOD3 is differentially localized to vitreous substructures and dysregulation of these interactions or SOD3 enzymatic activity may be critical in the pathophysiology of DVR. Project goals include: identifying SOD3 protein/proteoglycan interaction mechanisms in the human vitreous, determining the functional effects of SOD3 glycation in DVR, and investigating SOD3 oxidative stress pathways in the mouse and human vitreous. Several different techniques will be utilized. First, protein interactions will be assessed by co-immunoprecipitation, western blotting, and immunohistochemistry using native, recombinant, and mutant SOD3. Next, the functional effects of SOD3 glycation will be studied in vivo and in vitro. Finally, the balance of SOD3, reactive oxygen species, and oxidative damage will be measured in human and mouse DVR tissue samples using EPR spectroscopy, immunohistochemistry and western blotting. Detailed clinical phenotyping of SOD3 knockout mice will be performed and activation of oxidative stress pathways in the retina will be determined. The project is significant to human health because understanding SOD3-related oxidative stress pathways in the eye gives insight into the mechanisms of DVR pathophysiology and provides a unique therapeutic modality.