通过可控干燥系统建立兔干眼模型  

Rabbit Dry Eye Model Induced by a Controlled Drying System

作　　者：陈晓敏 邝健标 武振宁 周世有[1] CHEN Xiao-min;KUANG Jian-biao;WU Zhen-ning;ZHOU Shi-you(State Key Laboratory of Ophthalmology//Zhongshan Ophthalmic Center,Sun Yat-sen University,Guangzhou 510060,China;Lee′s Pharmaceutical Holdings Limited//Guangzhou Zhaoke Pharmaceutical Co.ltd,Guangzhou 511462,China)

机构地区：[1]中山大学中山眼科中心//眼科学国家重点实验室,广东广州510060 [2]李氏大药厂控股有限公司//兆科(广州)眼科药物有限公司,广东广州511462

出　　处：《中山大学学报（医学科学版）》2020年第2期251-259,共9页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(31671210)。

摘　　要：[目的]旨在通过可控干燥系统建立一种由干燥环境引起的兔干眼模型.[方法]本研究共使用24只正常雄性新西兰兔,随机分为对照组和干燥环境组,每组12只.干燥环境组在可控干燥系统(CDS)中,其相对湿度、空气流速和温度分别保持在(22±4)%、3~4 m/s和23℃~25℃,而对照组则饲养在普通环境中,相对湿度、空气流速和温度分别在60%~70%、0.2 m/s和23℃~25℃.两组动物饲养14 d.在实验的第0、3、7、14天进行Schirmer试验、角膜荧光素染色、结膜丽丝胺绿染色.第14天对动物施行安乐死,制备眼球组织切片,染色后通过光学显微镜评估角膜上皮厚度和结膜杯状细胞数量;使用免疫荧光染色方法检测结膜上皮中的MUC5AC蛋白表达水平;对眼表Caspase-3进行免疫组织化学染色,评估眼表凋亡水平.[结果]与对照组相比,在第3、7、14天干燥环境组出现泪液分泌减少(P<0.001),同时出现角膜荧光素染色增加和结膜丽丝胺绿染色增加(P<0.001).在第14天,对照组和干燥环境组的角膜上皮厚度分别为(58.0±7.2)μm和(47.8±7.6)μm,干燥环境组的角膜上皮厚度明显减低(P<0.05);结膜杯状细胞数量上对照组和干燥环境组分别为15±4和10±2,干燥环境组的数量明显减少(P<0.01);干燥环境组的结膜MUC5AC的表达量下降(与杯状细胞数量减少相一致);同时,干燥环境组的角膜上皮Caspase-3表达量明显增加,对照组和干燥环境组的积分光密度分别为(17±2)%和(20±2)%(P<0.01).[结论]暴露于干燥环境会导致兔眼表上皮出现干眼症体征及病理性改变.利用可控干燥系统构建的兔干眼模型将有望成为研究干眼症发病机理的新手段.【Objective】To provide a rabbit model for dry eye using a dry environment induced by Controlled Drying System.【Methods】Twenty-four male New Zealand rabbits were used in the experiment.They were randomly divided into control group and dry group,each one with 12 rabbits.The dry group was randomly housed in Controlled Drying System(CDS)for 14 days.The relative humidity,airflow and temperature were kept at(22±4)%,3~4 m/s and(23~25)℃,respectively.The control group were fed in a normal environment where relative humidity,airflow and temperature were kept at 60%~70%,0.2 m/s and(23~25)℃.The Schirmer test,corneal fluorescein staining,conjunctival lissamine green staining were performed during the experimental process on days 0,3,7,and 14.On the last day,the rabbits were euthanized and the eye tissues were made into paraffin-cut sections.After staining,we evaluated the corneal epithelial thickness and goblet cell number in the conjunctiva using light microscopy.MUC5AC in the conjunctival epithelium was detected by immunofluorescence.The apoptosis level changes on the ocular surface were evaluated using Caspase-3 by immunohistochemistry.【Results】Decreased tear production,increased corneal fluorescein staining and increased conjunctival lissamine green staining were found on days 3,7,and 14 in the dry group compared with the control group(P<0.001).Corneal epithelial thickness of control group and dry group were(58.0±7.2)μm and(47.8±7.6)μm,which showed corneal epithelial thickness of dry group was decreased(P<0.05).Goblet cells in the conjunctiva of control group and dry group were 15±4 and 10±2,which showed goblet cells of dry group was decreased(P<0.01).The expression of MUC5AC(consistent with goblet cells deficiency)was also reduced.Caspase-3 was highly expressed on the corneal epithelium in the dry group.IOD/field of control group and dry group were(17±2)%and(20±2)%(P<0.01).【Conclusions】Dry environment can make rabbits have pathological changes of dry eye on ocular surface epithelium.This dry e

关 键 词：干眼症 可控干燥系统 干燥环境 家兔 

分 类 号：R771.2[医药卫生—眼科]