非肥胖性糖尿病小鼠干眼模型的初步建立

Preliminary establishment of dry eye model in non-obese diabetic mice

目的:初步建立非肥胖性糖尿病(NOD)小鼠蒸发过强型干眼模型,通过研究小鼠眼表组织病理变化,初步探讨其作为糖尿病性干眼模型的可行性。方法:选取40只雌性NOD小鼠,NOD小鼠自发糖尿病为实验组,同时选取未自发糖尿病的NOD小鼠作为正常对照组。实验组NOD小鼠置于40%以下湿度环境,每天皮下注射0.5mg/0.2mL氢溴酸菪胺,并置于可控干燥箱中,每天通风12h,制作蒸发过强型干眼模型。在造模后的第1、7、10、14d采用酚红棉线实验测量泪液分泌量,PAS染色检查结膜杯状细胞形态和数目;在造模后的第10d,进行角膜组织苏木精染色检测角膜上皮变化情况。结果:实验组NOD小鼠泪液分泌量随造模时间而逐渐降低,正常对照组未有明显变化。实验组的结膜杯状细胞体积变大,在造模后的第1d,杯状细胞密度较正常对照组减少(P=0.008),从造模后第7d开始,随时间延长,实验组杯状细胞数量逐渐减少,且较同一时间点正常对照组显著减少(均P<0.001)。此外,观察两组第10d的角膜上皮情况,实验组NOD小鼠角膜上皮层变薄,部分角膜上皮细胞变性、基底细胞水肿。结论:初步建立了NOD小鼠干眼模型,其眼表变化与临床上干眼症表现类似。

AIM:To determine the pathological changes in ocular surfaces dry eye excessive evaporation non-obese diabetic (NOD) mice model and to preliminarily explore the feasibility of diabetic dry eye model. METHODS: In this study, 40 females NOD mice were selected. The experimental group consisted of NOD mice that were diagnosed with diabetes while the normal control group consisted of those NOD mice without spontaneous diabetes. Hypodermic injection of Scopolamine hydrobromide (0.5mg/0.2mL) was administered under 40% humidity to the experimental group and placed in a controlled drying box for 12h a day. This was to achieve a dry eye model. Testing indicators on the 1, 7, 10 and 14d after modeling, phenol red thread test was used to measure tear secretion and the eye sections were stained with periodic acid-Schiff (PAS) to examine the morphology and number of conjunctival goblet cells. On the 10d after modeling, the changes in the corneal epithelium were visualized after staining with hematoxylin. RESULTS:For the NOD mice of the experimental group, the tear secretion was gradually decreased with timing, while there were no obvious changes in the normal control group. The volume of the conjunctival goblet cells of the experimental group became larger, and on the 1d after the molding, the experimental group had decreased density of the goblet cells when compared with the normal control group ( P =0.008). From the 7d after the molding, as the time was prolonged, the density of the goblet cells was gradually decreased and the differences between the two group at same time point were significant (all P <0.001). Besides, it was required to observe the corneal epithelium of the two groups on the 10d. The result shows that the corneal epithelium became thinned, some epithelial cells were denatured, and stromal cells became edema. CONCLUSION: Dry eye model of NOD mice was preliminary established, and the changes of ocular surface were similar to those of dry eye in the clinic.