Jeong Jae Seung, Lee Woon-Kyu, Moon Yeon Sung, Kim Na Rae
Department of Ophthalmology and Inha Vision Science Laboratory, Inha University School of Medicine, Incheon, Korea.
Laboratory of Developmental Genetics, Department of Biomedical Sciences, Inha University School of Medicine, Incheon, Korea.
Lab Anim Res. 2017 Sep;33(3):216-222. doi: 10.5625/lar.2017.33.3.216. Epub 2017 Sep 27.
This study aims to evaluate early changes in retinal structure and BMP2 expression in the retina and crystalline lens by comparing streptozotocin-induced diabetic pigs and normal control group pigs.
Five eye samples from five diabetic Micro-pigs (Medikinetics, Pyeongtaek, Korea) and five eye samples from five control pigs bred in a specific pathogen-free area were used. Diabetes was developed through intravenous injection of nicotinamide and streptozotocin, and the average fasting glucose level was maintained at 250 mg/dL or higher for 16 weeks. To evaluate BMP2 expression in the retina and crystalline lens, Western blotting was performed.
In Hematoxylin and Eosin staining, most diabetic pigs showed structural abnormalities in the inner plexiform layer. The number of nuclei in the ganglion cell layer within the range of 10 µm was 3.78±0.60 for diabetic pigs and 5.57±1.07 for control group pigs, showing a statistically significant difference. In immunohistochemical staining, diabetic retinas showed an overall increase in BMP2 expression. In Western blotting, the average BMP2/actin level of diabetic retinas was 1.19±0.05, showing a significant increase compared to the 1.06±0.03 of the control group retinas (=0.016). The BMP2/actin level of diabetic crystalline lenses was similar to the control group crystalline lenses (=0.730).
Compared to control group pigs, the number of nuclei in the inner nuclear layer of retinas from streptozotocin-induced diabetic pigs decreased, while an increase in BMP2 expression was observed in the retina of diabetic pigs.
本研究旨在通过比较链脲佐菌素诱导的糖尿病猪和正常对照组猪,评估视网膜结构以及视网膜和晶状体中骨形态发生蛋白2(BMP2)表达的早期变化。
使用来自5只糖尿病微型猪(韩国平泽市Medikinetics公司)的5只眼样本和来自在无特定病原体区域饲养的5只对照猪的5只眼样本。通过静脉注射烟酰胺和链脲佐菌素诱导糖尿病,平均空腹血糖水平维持在250mg/dL或更高达16周。为评估视网膜和晶状体中BMP2的表达,进行了蛋白质免疫印迹分析。
苏木精-伊红染色显示,大多数糖尿病猪的内网状层存在结构异常。糖尿病猪在10μm范围内神经节细胞层的细胞核数量为3.78±0.60,对照组猪为5.57±1.07,差异具有统计学意义。免疫组织化学染色显示,糖尿病视网膜中BMP2表达总体增加。蛋白质免疫印迹分析显示,糖尿病视网膜的平均BMP2/肌动蛋白水平为1.19±0.05,与对照组视网膜的1.06±0.03相比显著增加(P=0.016)。糖尿病晶状体的BMP2/肌动蛋白水平与对照组晶状体相似(P=0.730)。
与对照组猪相比,链脲佐菌素诱导的糖尿病猪视网膜内核层的细胞核数量减少,而糖尿病猪视网膜中BMP2表达增加。