建立在体角膜内皮细胞损伤后再生动物模型的实验研究  

Experimental study on regeneration of corneal endothelial cells in vivo

作　　者：苏冠羽 韦振宇 王乐滢 梁庆丰 Su Guanyu;Wei Zhenyu;Wang Leying;Liang Qingfeng(Beijing Tongren Eye Center,Beijing Tongren Hospital,Capital Medical University,Beijing Institute of Ophthalmology,Beijing Key Laboratory of Ophthalmology and Visual Sciences,Beijing,100005,China)

机构地区：[1]首都医科大学附属北京同仁医院北京同仁眼科中心北京市眼科研究所眼科学与视觉科学北京市重点实验室

出　　处：《中华眼科医学杂志（电子版）》2019年第5期298-304,共7页Chinese Journal of Ophthalmologic Medicine(Electronic Edition)

基　　金：国家自然科学基金面上项目(81470607);北京市卫生系统高层次卫生技术人才培养基金(2014-3-016);2017年北京市百千万人才工程培养项目(2017A10)

摘　　要：目的探讨建立在体角膜内皮损伤动物模型的方法及其损伤后再生的潜能。方法选取2周龄的正常六角龙鱼30只。采用数字表法随机分为正常观察组、NaOH损伤组及机械损伤组,各10只。采用活体共聚焦显微镜检查、组织病理学检查、茜素红-曲利苯蓝内皮染色和氯化金角膜神经染色等方法观察。对正常观察组六角龙鱼,观察其眼球和角膜正常结构;通过NaoH溶液和器械两种方法损伤角膜内皮层,建立角膜内皮细胞损伤模型,观察其角膜内皮细胞再生的情况。对4个时间点角膜内皮细胞的密度进行正态性检验和方差齐性检验,使用重复测量方差分析对不同组别、不同时间点角膜内皮细胞密度的均值进行比较,事前进行Mauchly球形检验,如果检验结果显著,采用多元方差分析,否则选用校正自由的F检验。结果六角龙鱼角膜厚度约为(75.75±7.51)μm,角膜上皮细胞层较厚,约占角膜厚度的一半。经NaoH溶液和器械两种方法损伤其角膜内皮细胞后,六角龙鱼角膜内皮细胞密度明显降低。NaoH损伤组和机械损伤组在不同时间点,六角龙鱼角膜内皮细胞密度的比较,具有统计学意义(F=31.38,51.77;P<0.05)。而各个时间点间的差异,两组均无统计学意义(F=1.37,2.67,0.70,4.14;P>0.05)。在损伤后3 d时,NaoH损伤组六角龙鱼角膜内皮细胞的密度为(128±14)个/mm^2,机械损伤组为(113±11)个/mm^2。与损伤前比较,其角膜内皮细胞密度的差异均有统计学意义(t=19.39,8.78;P<0.05);在损伤后7 d时,NaoH损伤组六角龙鱼角膜内皮细胞的密度为(157±20)个/mm^2,机械损伤组为(169±19)个/mm^2。与损伤后3d时比较,其角膜内皮细胞密度均有所恢复,差异均有统计学意义(t=3.75,8.07;P<0.05);在损伤后14 d时,可见NaoH损伤组六角龙鱼角膜内皮细胞的密度为(198±17)个/mm^2,机械损伤组为(223±17)个/mm^2。与损伤后3 d时和7 d时比较,均有明显恢复,差异均有统计学�Objective This study was aim to explore the method of establishing animal model of corneal endothelial injury and the potential of endothelial regeneration after injury. Methods 30 normal axolotls of 2 weeks old were selected. They were divided into normal observation group, NaoH-injured group and mechanical-injured group with 10 axolotls each. In vivo confocal microscopy, histopathological examination, alizarin red-trichlorobenzene blue endothelial staining and gold chloride corneal nerve staining were used to observe. In the normal observation group, the normal structure of the eye and cornea was observed. Corneal endothelial cells were damaged by NaoH solution and mechanics, and the endothelial cell injury model was established to observe the regeneration of corneal endothelial cells. The corneal endothelial cell density at four time points was tested by normality test and homogeneity test of variance. The mean corneal endothelial cell density at different time points in different groups was compared by repeated measurement variance analysis. Mauchly spherical test was performed beforehand. According to the test results,if significant,the results of multivariate analysis of variance are adopted;otherwise,the correction free F test is selected. Results The corneal thickness of axolotls was about( 75. 75 ± 7. 51) μm,and the corneal epithelial cell layer was about half of the corneal thickness. After the corneal endothelial cells were damaged by Nao H solution and mechanic,the density of corneal endothelial cells decreased significantly.The density of corneal endothelial cells in Nao H-injured group and mechanical-injured group at different time points was significantly different( F = 31. 38,51. 77;P < 0. 05). There was no significant difference between Nao H-injured group and mechanical-injured group at each time point( t = 1. 37,2. 67,0. 70,4. 14;P > 0. 05). At 3 days after injury,the density of corneal endothelial cells in Nao H-injured group and mechanical-injured group were( 128 ± 14)/mm2 and( 113 ± 11)/

关 键 词：动物模型 角膜 组织病理 内皮损伤 

分 类 号：R-332[医药卫生] R772.2