Mitochondrial Superoxide Anion Overproduction in Tet-mev-1 Transgenic Mice Accelerates Age-Dependent Corneal Cell Dysfunctions
Mitochondrial Superoxide Anion Overproduction in Tet-mev-1 Transgenic Mice Accelerates Age-Dependent Corneal Cell Dysfunctions
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DOI：
10.1167/iovs.12-9573
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发表时间：
2012-08-01
影响因子：
4.4
通讯作者：
Ishii, Naoaki
中科院分区：
文献类型：
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作者：
Onouchi, Hiromi;Ishii, Takamasa;Ishii, Naoaki
目的.表达由四环素（泰特）-开/关系统控制的线粒体复合物-II突变的SDHCV 69 E基因的Tet-mev-1小鼠可以过量产生O-2（中心点-），并且是用于研究线粒体氧化应激的通用整体动物模型。在此，我们报道了Tet-mev-1小鼠与野生型C57 BL/6 j小鼠相比，角膜上皮、内皮和实质细胞的一系列年龄依赖性变化。测定（1）线粒体电子传递酶活性;（2）O-2（中心点）产生;（3）作为氧化应激标志物的羰基化蛋白和8-羟基脱氧鸟苷（8-OHdG）水平;（4）光学和电子显微镜下的病理学分析;（5）苏木精-伊红或甲苯胺蓝染色;和（6）用抗-β-连环蛋白抗体进行免疫组织化学，在眼睛中，特别是在角膜中。复合物II和辅酶Q之间的电子泄漏降低了复合物II-III的活性。这导致Tet-mev-1小鼠眼睛中年龄依赖性细胞内氧化应激增加。20%乙醇处理后，Tet-mev-1小鼠角膜上皮形成延迟，因为与野生型相比，Tet-mev-1小鼠角膜上皮中的细胞和有丝分裂细胞数量减少。角膜内皮细胞中细胞数量的年龄依赖性减少加速。此外，Tet-mev-1小鼠角膜厚度随着年龄的增长而减少，其实质细胞变薄。这些结果表明，线粒体氧化应激与电子传递链功能障碍可以影响年龄相关性角膜疾病的发病机制和进展，以及广义的角膜老化加速。（Invest Ophthalmol维斯科学。2012;53：5780-5787）DOI：10.1167/iovs.12-9573
PURPOSE. The Tet-mev-1 mouse expressing a mitochondrial complex-II mutated SDHCV69E gene controlled by a tetracycline (Tet)-On/Off system can overproduce O-2(center dot-) and is a versatile whole-animal model for studying mitochondrial oxidative stress. Here we report a series of age-dependent variations in corneal epithelium, endothelium, and parenchymal cells of the Tet-mev-1 mice relative to wild-type C57BL/6j mice.METHODS. Measurements of (1) mitochondrial electron transport enzyme activities; (2) O-2(center dot-) production; (3) carbonylated protein, and 8-hydroxydeoxyguanosine (8-OHdG) levels as markers of oxidative stress; (4) pathologic analyses under optical and electron microscopy; (5) hematoxylin-eosin or toluidine-blue staining; and (6) immunohistochemistry with an anti-beta-catenin antibody were performed in the eye, especially the cornea.RESULTS. Complex II-III activity was decreased by electron leakage between complex II and CoQ. This resulted in increased age-dependent intracellular oxidative stress in the eye of Tet-mev-1 mice. Corneal epithelialization was delayed in Tet-mev-1 mice after 20% ethanol treatment, as the number of cells and mitotic cells decreased in the corneal epithelium of Tet-mev-1 mice compared with that of wild type. The age-dependent decrease in cell number accelerated in the corneal endothelium cells. Moreover, it was suggested that the corneal thickness was decreased by thinning of parenchymal cells with age in Tet-mev-1 mice.CONCLUSIONS. These results suggest that mitochondrial oxidative stress with electron transport chain dysfunction can influence pathogenesis and progression of age-related corneal diseases, as well as generalized corneal aging acceleration. (Invest Ophthalmol Vis Sci. 2012;53:5780-5787) DOI:10.1167/iovs.12-9573