异硫氰酸荧光素-葡聚糖眶后注射观察鼠视网膜血管的可行性研究

摘要

背景 糖尿病视网膜病变、早产儿视网膜病变、年龄相关性黄斑变性等视网膜血管性疾病的发病率逐年提高,客观有效地评价视网膜的血管情况是视网膜血管性疾病防治研究的关键.异硫氰酸荧光素-葡聚糖(FITC-dextran)眶后注射法是评价C57BL/6J小鼠视网膜血管的新方法,但目前少有关于此方法是否适合于其他小鼠及大鼠研究的报道. 目的 评估用FITC-dextran眶后注射法观察实验常用鼠种视网膜血管的可行性,为相关的实验研究提供参考依据.方法 选择SPF级C57BL/6J小鼠、昆明小鼠、SD大鼠、Wistar大鼠各12只,均分为实验组和对照组各6只.实验组动物右眼眶后注射9 ml/kg FITC-dextran溶液,对照组右眼眶后注射等体积PBS溶液.注射10 s后大鼠以过量麻醉法处死,小鼠以颈椎脱臼法处死,摘取双侧眼球,荧光显微镜下行双侧眼球后组织以及视网膜铺片检查.结果 C57BL/6J小鼠、昆明小鼠实验组双眼均可以观察到FITC-dextran绿色荧光标识的视网膜血管,但对照组各眼均观察不到视网膜血管形态；SD大鼠、Wistar大鼠实验组及对照组受检眼在荧光显微镜下均未观察到FITC-dextran标识的视网膜血管.小鼠、大鼠实验组右眼均可见由FITC-dextran浸染的绿色球后组织,而左眼球后组织均未见荧光. 结论 FITC-dextran眶后注射法适合于观察小鼠的视网膜血管,不适于观察大鼠的视网膜血管.%Background The incidence of retinal vascular diseases increase annually,such as diabetic retinopathy,retinopathy of prematurity and age-related macular degeneration.The key of treatment for these diseases is how to evaluate retinal vascular change effectively and objectively.Retro-orbital injection of fluorescein isothiocyanatedextran (FITC-dextran) is a simple and effective method for observing C57BL/6J mouse retinal vessels.But,whether it is suitable for other mice and rats is seldom reported.Objective This experiment was to assess the feasibility of the observation of retinal vessels by retro-orbital injection of FITC-dextran in different genus of mouse and offer the reference for relevant study.Methods Twelve animals of C57BL/6J mice,Kunming mice,SD rats and Wistar rats were selected,respectively and divided into the experimental group and control group at average.The right eyes of the animals of the experimental group received the retro-orbital injection of 9 ml/kg FITC-dextran,and the right eyes of animals of the control group received PBS solution at the same volume and way.All the animals were sacrificed 10 seconds after injection and both eyes of each animal were obtained for retinal stretched preparation.The retrobulbar tissue and whole-mount retina were viewed under a fluorescence microscope.The use of the animals complied with Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results Retinal blood vessels labeled by FITC-dextran could be observed in both eyes of C57BL/6J mice and Kunming mice to present with a green fluorescence in experimental group under a fluorescence microscope,but no any fluorescence-labeled retinal blood vessel was exhibited in the control mice.The retinal blood vessel could not be observed in all eyes of SD rats and Wistar rats after the injection of FITC-dextran both in the experimental group and the control group under a fluorescence microscope.The surrounding tissues of the right eyes of mice and rats dyed with green fluorescence of FITC-dextran in the experimental group,however,green fluorescence could not be seen in the surrounding tissues of the left eyes of mice and rats.Conclusions Retro-orbital injection of FITC-dextran is a suitable method of observing the retinal vessels of mouse but not rat.