慢性高眼压青光眼动物模型的构建和鉴定  

Establishment and appraisal of glaucoma animal models with chronic intraocular hypertension

作　　者：马建洲[1] 贺翔鸽[1] 谢琳[1] 许建涛[1] 孙亚丽[2] 王永堂[3] 龙在云[3] 李应玉[3] 

机构地区：[1]第三军医大学大坪医院野战外科研究所眼科,中国重庆市400042 [2]烟台毓璜顶医院病理科,中国山东省烟台市264000 [3]第三军医大学大坪医院野战外科研究所3室,中国重庆市400042

出　　处：《国际眼科杂志》2007年第4期951-955,共5页International Eye Science

摘　　要：目的:建立大鼠慢性高眼压模型,观察眼压、病理和视功能改变。方法:SD大鼠45只,麻醉后使用532二极管激光行右眼角膜缘360°光凝,角膜缘激光斑为80～100个,大鼠角膜缘颞侧、颞上及颞下巩膜浅层静脉3条,每条静脉光凝3～4个斑点,功率0.45W/0.7s。左眼为对照眼,3d后测量眼压,部分眼压升高不明显者,进行同样的二次光凝。Tono-penXL眼压计监测3,7,30,60,90,180d麻醉状态下的眼压。激光术后60,180d取大鼠各5只,40g/L多聚甲醛灌注固定,摘取双侧眼球和视神经,分别进行冰冻和石蜡切片,采用HE染色、尼氏染色、甲苯胺蓝染色,光镜下观察房角变化,不同时间视网膜节细胞计数,比较视神经髓鞘密度的变化。60,180d大鼠10只,使用TEC-350V视觉电生理仪行F-VEP检查;然后6mo大鼠5只,进行逆行荧光金标记RGCs,7d后40g/L多聚甲醛灌注固定,全视网膜铺片,荧光显微镜下比较视网膜节细胞数量变化。结果:大鼠高眼压模型成功38只,平均最高眼压在激光后30d,平均值为25.0±4.1mmHg,对照眼17.1±3.2mmHg,180d时眼压基本恢复正常。病理改变:实验眼前房角明显变窄,小梁网间隙压缩、变窄,甚至部分闭锁,消失,少量梭形成纤维细胞聚集,组织致密、硬化,而小梁细胞减少,虹膜部分卷曲,水肿、肥厚出现明显异常。逆行荧光金视网膜铺片和视网膜切片尼氏染色见实验眼视网膜节细胞数量有明显的减少;尼氏染色切片每400倍视野总平均数,60d组对照眼为41±10.6个,实验眼为35±11.2个,180d组对照眼为40±9.8个,实验眼为34±11.0个,周边视网膜平均值减少最为显著,均值相差可达8个神经节细胞;180d时模型眼视神经甲苯胺蓝染色显示的髓鞘密度明显降低;视功能检查:高眼压大鼠模型60,180d的实验眼和对照眼均可引出典型的和重复性好的NPN波形,60d时实验眼AP1(N1-P1振幅)均值降低,为13.03±3.11ms,对照眼为21.14±3.10ms,两者具有显著性差异(P�AIM： To develop a distinctive model of the chronic glaucoma with optic neuropathy induced by intraocular hypertension in rats. METHODS： The rat model with chronic elevation of intraocular pressure （IOP） was established and the changes of optic nearopathy were observed. A diode laser with wavelength of 532-nm was used to burn the trabecular meshwork and episcleral veins of right eyes at 45 SD rats, while the left eyes were referred as control. Tono-pen XL Tonometer was used to measure IOP at 3 days; 1 week;1,2 3, 6 months under Pentobarbital Sodium anesthesia. The F-VEP was monitored at the model of rats in 2 and 6 months after laser burning. The retina and optic nerve samples were obtained for morphological observation to judge whether the animal model was successful. RESULTS： 38 rat models with intraocular hypertension were obtained successfully. The IOP of experimental eyes in the morning was maintained at 25.0±4.1mmHg in the first month. It was increased in all eyes to higher than the control eyes, the mean IOP of which was 17.1±3.2mmHg, and then it decreased after 3 months. The mean IOP was the same as that in the control in 6 months. CONCLUSION： The rat model with chronic elevation of IOP caused by a laser injury to the trabecular meshwork represents a useful and efficient experiment model of rats with glaucoma.

关 键 词：青光眼 视神经 视网膜 大鼠 免疫荧光染色 激光光凝 

分 类 号：R775[医药卫生—眼科]