Development of Animal Models for Lens and Corneal Diseases Using N-Methyl-N-Nitrosourea
Development of Animal Models for Lens and Corneal Diseases Using N-Methyl-N-Nitrosourea
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使用 N-甲基-N-亚硝基脲开发晶状体和角膜疾病动物模型
DOI：
10.1167/iovs.61.8.38
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发表时间：
2020-07-01
影响因子：
4.4
通讯作者：
Huang, Yifei
中科院分区：
文献类型：
--
作者：
Qu, Yingxin;Li, Runpu;Huang, Yifei
目的N-甲基-N-亚硝基脲（MNU）是一种具有强致突变性的烷化剂。本研究旨在通过MNU给药诱导透镜上皮细胞（LEC）和角膜内皮细胞（CEC）凋亡。我们试图建立白内障和角膜内皮失代偿的眼病模型。方法将MNU注入新生大鼠腹腔和成年家兔前房。然后在不同时间点对MNU处理的动物进行一系列功能和形态学分析。结果MNU处理可诱导LEC和CEC广泛凋亡。这些效应具有剂量和时间依赖性。MNU处理后3周，在新生大鼠中发现成熟的白内障。组织学分析表明，MNU毒性诱导肿胀，空泡化，液化的透镜纤维的MNU治疗的大鼠。Pentacam检查显示MNU给药后大鼠透镜的平均密度显著增加。末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）分析显示，在MNU治疗的大鼠晶状体中普遍存在凋亡染色。在兔眼中，前房内注射MNU诱导角膜水肿，并显著增加中央角膜厚度，在P14达到峰值。形态学和免疫组织化学分析表明，CEC有效地消融在MNU治疗的兔子。8-OHdG的表达在MNU处理的兔角膜中显著增加，与溶剂处理的对照组相比。结论在动物模型中，MNU足以诱导眼细胞凋亡。MNU诱导的白内障和角膜内皮失代偿的这些模型代表了用于开发相关疗法的有价值的工具。
Purpose N-methyl-N-nitrosourea (MNU) is an alkylating toxicant with potent mutagenic ability. This study was designed to induce apoptosis in lens epithelial cells (LECs) and corneal endothelial cells (CECs) via MNU administration. We sought to build ocular disease models of cataract and corneal endothelial decompensation. Methods MNU was delivered into the intraperitoneal cavities of neonatal rats and the anterior chambers of adult rabbits. The MNU-treated animals were then subjected to a series of functional and morphological analyses at various time points. Results MNU treatment induced pervasive apoptosis of LECs and CECs. These effects were dose and time dependent. Mature cataracts were found in neonatal rats 3 weeks after MNU treatment. Histological analysis revealed that MNU toxicity induced swelling, vacuolation, and liquefaction in lens fibers of MNU-treated rats. Pentacam examination showed that the average density of rat lens increased significantly after MNU administration. Terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) analysis showed pervasive apoptotic staining in the lenses of MNU-treated rats. In rabbit eyes, intracameral treatment with MNU induced corneal edema and significantly increased central corneal thickness, which peaked at P14. Morphological and immunohistochemical analysis showed that CECs were effectively ablated in the MNU-treated rabbits. The expression of 8-OHdG increased significantly in the cornea of MNU-treated rabbits, compared with vehicle-treated controls. Conclusions MNU is sufficient to induce ocular cell apoptosis in animal models. These models of MNU-induced cataract and corneal endothelial decompensation represent valuable tools for efforts to develop relevant therapies.