小剂量多次注射链脲佐菌素建立糖尿病早期视网膜病变动物模型  

Establishment of a rat model of early diabetic retinopathy induced by multiple low-dose streptozotocin injection

作　　者：朱华[1] 李彦红[1] 徐艳峰[1] 徐玉环[1] 许庆刚[2] 尹红霞[2] 秦川[1] 

机构地区：[1]卫生部人类疾病比较医学重点实验室,国家中医药管理局人类疾病动物模型三级实验室,中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,北京100021 [2]首都医科大学附属同仁医院,北京100730

出　　处：《中国实验动物学报》2016年第5期487-493,共7页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金青年项目(编号:81201075);国家自然基金面上项目(编号:81371546)

摘　　要：目的多次小剂量注射链脲佐菌素(STZ)制作糖尿病(DM)早期大鼠视网膜病变(DR)动物模型。方法雄性SD大鼠75只,体重(180±15)g。随机分为对照组(CON组,n=30)和模型组(DM组,n=45)。DM组按30mg/kg体重腹腔注射3%的STZ,连续5 d。成模后每周测量体重、血糖等指标。分别在成模后第4、8、12周各组随机安乐死10只动物,对视网膜组织进行H.E染色、免疫组化、消化铺片及透射电镜超微结构观察。结果模型成立后第2周,动物表现出多食、多饮、多尿的糖尿病症状。与对照组比较,模型大鼠4周时仅见视网膜变薄,8周时视网膜内皮细胞增生,毛细血管基底膜增厚,内皮细胞指状突起,吞饮泡增多。12周时毛细血管膨大,毛细血管细胞凋亡,核固缩、深染。视网膜变薄,神经节细胞数量减少。视网膜周细胞线粒体肿胀,嵴脱落空泡样变。模型组大鼠视网膜GFAP阳性细胞主要分布在节细胞层和神经纤维层,呈条索状连续排列,阳性细胞数明显减少。结论小剂量多次注射STZ诱导的大鼠DR模型表现出人类早期视网膜病变相似的特征,可用于发病机理、药效学等方面的研究。Objective The aim of this study was to observe the morphological changes of the retina at early stage of diabetes in SD rat induced by multiple injections of low-dose（ multi-low-dose） streptozotocin（ STZ）. Methods Seventy-five male SD rats were randomly divided into control（ CON）（ n = 30） and diabetes mellitus（ DM）（ n = 45） groups.Rats in the DM group received multiple low-dose STZ i. v. injection for 5 consecutive days（ 30 mg/kg）. The blood glucose,body weight,and other parameters were observed once a week after injection. In the 4,8 and 12 w,the morphological changes in the retina were observed using HE and immunohistochemical（ IHC） staining,vasculature digest preparation and transmission electron microscopy（ TEM）. Results The body weight of animals in the DM group was decreased from 2w. The average body weight of the DM group was significantly lower than that of CON group（ P〈0. 01）. The average blood glucose of DM group was significantly higher than that of the CON group（ P〈0. 01）. TEM examination revealed thickened capillary basement membrane and dilatation of capillary at 4 w. trypsin digest preparation showed endothelial cell hyperplasia,mitochondrial swelling,cristae disruption,and vacuolar degeneration in capillary endothelial cells at 8 w.Retina HE staining showed retinal capillary dilatation and interstitial edema,capillary intumescence,apoptosis in endothelial cells in the DM group at 12 w. The control group had no obvious abnormalities. Staining of GAFP in the retina indicated decreased expression in the ganglion cell layer and nerve fiber layer,bipolar cells and ganglion cells,mitochondrial swelling and cristae disruption in pericytes,decreased membranous disc,and widened gap between membranous discs at 12 w. Conclusions Our findings indicate that an early stage diabetic retinopathy（ DR） can be induced in rats by multiple low-dose streptozotocin injection. This may serve as a valuable preclinical model for studying the pathogenesis,pharmacodynamics

关 键 词：糖尿病 糖尿病视网膜病 动物模型 链脲佐菌素 大鼠 

分 类 号：Q95-33[生物学—动物学]