摘要
目的:探讨小鼠干眼模型构建及其角膜特征检测方法。方法:选取健康无特定病原体(SPF)级C57BL/6小鼠共12只，6~8周龄，雄性，体重(20±1)g。采用数字表法将12只小鼠随机分为实验组和对照组，每组6只，选择每只小鼠的右眼用于实验。实验组6只小鼠(6只眼)右眼给予0.2%苯扎氯铵溶液5 μL局部点眼构建小鼠干眼模型，每天9：00和21：00点眼，共2次，持续7 d；对照组6只小鼠(6只眼)右眼给予磷酸盐缓冲溶液5 μL局部点眼作为正常对照，每天9：00和21：00点眼，共2次，持续7 d。在7 d时，采用动物眼科裂隙灯显微镜对实验组及对照组小鼠右眼行角膜荧光素染色检查，并进行角膜荧光素染色评分；应用ISOCT眼科超显微成像系统对实验组和对照组小鼠右眼行眼前节光学相干断层扫描(AS－OCT)检查，测量并记录角膜中央上皮层厚度。角膜荧光染色评分及角膜中央上皮层厚度符合正态分布，以(±s)进行描述，采用非配对t检验比较两组之间的差异。结果:在应用0.2%苯扎氯铵溶液造模后7 d，角膜荧光素染色成像结果显示实验组小鼠右眼角膜透明度下降且可见点片状着染，而正常对照组角膜透明，未见明显异常改变。实验组小鼠右眼角膜荧光素染色评分均较对照组增加，分别为(10.833±0.98)分和(1.833±0.98)分，其差异具有统计学意义(t＝15.85；P<0.05)。AS－OCT检查结果显示实验组小鼠右眼角膜上皮变形、厚度缺陷并呈现高反射点状角膜上皮浸润，实验组小鼠右眼角膜中央上皮层厚度较对照组明显下降，分别为(33.00±6.64)μm和(44.45±4.34)μm，其差异具有统计学意义(t＝3.53 ；P<0.05)。结论:0.2%苯扎氯铵溶液诱导的小鼠干眼模型相对稳定，AS－OCT适用于对苯扎氯铵诱导的小鼠干眼模型特征的检测，其特征主要表现为角膜上皮变形、厚度缺陷、呈高反射点状上皮浸润并存在明显的角膜中央上皮层厚度变薄。
Abstract
Objective:To explore the construction of a mouse dry eye model and its corneal feature detection method.Methods:A total of 12 healthy pathogen free (SPF) grade C57BL/6 mice, aged 6 to 8 weeks, male, weighing (20±1) g were selected. Using the number table method, 12 mice were randomly divided into an experimental group and a control group, with 6 mice in each group. The right eye of each mouse was selected for the experiment. Six mice (6 eyes) in the experimental group were given a 0.2% benzalkonium chloride solution 5 μL in their right eyes to construct a mouse dry eye model with local eye drops, twice a day at 9: 00 and 21: 00, lasting for 7 days; 6 mice in the control group (6 eyes) were given phosphate buffer solution 5 μ L in the right eye local eye drops were used as a normal control, with eye drops taken twice daily at 9: 00 and 21: 00, lasting for 7 days. At 7 days, corneal fluorescein staining was performed on the right eye of mice in the experimental and control groups using an animal ophthalmic slit lamp microscope, and corneal fluorescein staining scores were performed. The ISOCT ophthalmic ultra microscopic imaging system was used to perform optical coherence tomography (AS-OCT) examination of the anterior segment of the right eye of experimental and control group mice, and the thickness of the central corneal epithelial layer was measured. The corneal fluorescence staining score and the thickness of the central corneal epithelial layer conform to the normal distribution, and were described by (±s), and compared by the unpaired t test.Results:Seven days after modeling with 0.2% benzalkonium chloride solution, the results of corneal fluorescein staining imaging showed that the transparency of the right eye cornea of the experimental group mice decreased and spot staining, while the cornea of the normal control group was transparent, without obvious abnormal changes. The fluorescein staining score of the right eye cornea of the experimental group mice increased compared with the control group, which were (10.833±0.98) points and (1.833±0.98) points, respectively, with a statistically significant difference (t=15.85, P<0.05). AS-OCT examination results showed that the right corneal epithelium of mice in the experimental group was deformed, with thickness defects and high reflection punctate corneal epithelial infiltration. The thickness of the central corneal epithelial layer of mice in the experimental group was significantly lower than that in the control group, which were (33.00±6.64) μ m and (44.45±4.34) μm, respectively. The difference was statistically significant (t=3.53, P<0.05).Conclusions:The mouse dry eye model induced by 0.2% benzalkonium chloride solution is relatively stable, and AS-OCT is suitable for detecting the characteristics of the benzalkonium chloride induced mouse dry eye model. Its main features are corneal epithelial deformation, thickness defects, high reflex point like epithelial infiltration, and significant thinning of the central corneal epithelial layer.
基金项目
北京市百千万人才工程项目(2020027)
202国家自然科学基金青年项目(82101138)
万方数据