| 茅佩瑶,茅希颖,陈 雪,袁松涛,刘庆淮.碘酸钠诱导小鼠视网膜色素上皮变性模型的研究[J].现代生物医学进展英文版,2020,(9):1636-1641. |
| 碘酸钠诱导小鼠视网膜色素上皮变性模型的研究 |
| Study on a Mouse Model of Retinitis Pigmentosa Induced by Sodium Iodate |
| Received:February 20, 2020 Revised:March 15, 2020 |
| DOI：10.13241/j.cnki.pmb.2020.09.007 |
| 中文关键词: 视网膜色素变性 碘酸钠 视网膜色素上皮细胞 光感受器细胞 |
| 英文关键词: Retinitis pigmentosa Sodium iodate RPE Photoreceptor |
| 基金项目:国家自然科学基金项目（81970821）；国家重大科技专项（2017YFA0104101） |
| Author Name | Affiliation | E-mail | | MAO Pei-yao | Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210029, China | m9p2y0@163.com | | MAO Xi-ying | Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210029, China | | | CHEN Xue | Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210029, China | | | YUAN Song-tao | Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210029, China | | | LIU Qing-huai | Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210029, China | |
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| 中文摘要: |
| 摘要 目的：探讨股静脉注射不同剂量的碘酸钠后观察C57BL/6J小鼠视网膜色素上皮形态学及视网膜功能的变化。方法：选取30只6-8周龄C57BL/6J的雄性小鼠，随机分为正常对照组6只，实验1组（静脉注射碘酸钠10 mg/kg）6只，实验2组（静脉注射碘酸钠20 mg/kg）6只，实验3组（静脉注射碘酸钠35 mg/kg）6只，实验4组（静脉注射碘酸钠50 mg/kg）；正常对照组注射同等剂量的生理盐水。实验组分别经股静脉注射碘酸钠10、20、35、50 mg/kg，于注射后1周行电生理检测，1周、2周行OCT检测。所有小鼠2周后摘除眼球制作冰冻切片以及RPE细胞平铺片进行HE染色、免疫荧光染色。结果：正常对照组小鼠视网膜各层排列整齐，各层之间分界清晰，外核层形态正常，RPE层细胞排列紧密。注射后1周，通过ERG可发现碘酸钠10 mg/kg组小鼠视锥细胞功能已出现损伤，但OCT显示视网膜形态正常；而20 mg/kg以及35 mg/kg小鼠除了视锥细胞功能出现损伤，视杆细胞的功能均降至对照组1/2；且视网膜外核层均出现异常高亮区，外层视网膜丧失分界清晰的结构。注射后2周，10 mg/kg组小鼠通过RPE细胞平铺片即可发现RPE细胞已出现轻微损伤。而20 mg/kg、35 mg/kg组小鼠通过OCT可发现外核层与对照组相比均明显变薄，视网膜出现退行性改变；且通过HE染色和RPE细胞平铺片以及冰冻切片，我们可以发现20 mg/kg、35 mg/kg小鼠外核层出现波浪状改变，单层RPE细胞的连续性被破坏，呈现剂量依赖性。结论：碘酸钠20 mg/kg组经静脉注射后，可以很好的模拟年龄相关性黄斑变性的发病过程，视网膜出现明显的形态和功能变化，为视网膜色素变性提供一个较好的小鼠动物模型。 |
| 英文摘要: |
| ABSTRACT Objective: To characterize the efficiency of administrating different doses of sodium iodate by intravenous injection in imitating retinitis pigmentosa. Methods: Thirty 6-8 weeks old C57BL/6J mice was randomly divided into five groups: control group, 10 mg/kg NaIO3 group, 20 mg/kg NaIO3 group, 35 mg/kg NaIO3 group, 50 mg/kg NaIO3 group. Then different levels of sodium iodate were administered by intravenous injection into corresponding group of mice. Control group mice were given equivalent dose of PBS by intravenous injection. Functional changes of the retina were assessed at 1 week after injection by electroretinogram (ERG); morphological changes of the retina were assessed at 1,2 weeks following injection by optical coherence tomography (OCT). Eyeballs from all mice were removed 2 weeks after injection for frozen section and RPE flat mount, then HE is staining and immunostaining. Results: At 1 week after injection the amplitude of b wave from ectopic ERG in the 10 mg/kg group indicated a mild damage to function of cones, but morphologically the structure of ONL was normal; the amplitudes of b waves from no matter ectopic or scotopic ERG all underwent a drastic decline in the 20 mg/kg and 35 mg/kg group, which showing a damage both of cones but also of rods. Moreover, outer nuclear layer (ONL) of OCT results exerted higher reflection image than control, lacking distinctly stratified structure. At 2 weeks after injection, flat mounts of RPE from 10 mg/kg group exerted multinuclear cell which is a marker of RPE damage. In the 20 mg/kg and 35 mg/kg group, at 2 weeks after injection, the whole thickness of retina was decreased compared to control in a dose-dependent manner, which indicating a degeneration change to the retinas; moreover, H-E staining and immunostaining of RPE flat mount showed ONL in 20 mg/kg and 35 mg/kg groups were wave-like and continuity of monolayer RPE was interrupted, black round deposits and multinuclear RPE appearing. Immunostaining results of RPE specific marker RPE65 were declined both in 20 mg/kg and 35 mg/kg. Conclusion: The intravenous injection of sodium iodate can simulate the pathogenesis of retinitis pigmentosa in 20 mg/kg group, which can provide an animal model for retinitis pigmentosa. |
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