慢性高眼压大鼠模型视网膜中Brn-3a表达的动态变化  

The dynamic change of Brn-3a expression in retina of chronic high intraocular pressure rat

作　　者：朱益华[1] 陈晓莉[1] 

机构地区：[1]福建医科大学附属第一医院眼科,福州350005

出　　处：《中华实验眼科杂志》2011年第12期1077-1081,共5页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目（30471867）、福建省自然科学基金项目（C0520003）

摘　　要：背景Brn-3a是最近发现的一种视网膜神经节细胞（RGCs）特异性标记物。高眼压的视功能损害主要与RGCs损伤有关，但高眼压大鼠视网膜损伤与Brn-3a表达的关系尚不清楚。目的观察慢性高眼压大鼠不同时期眼压、视网膜的形态学和Brn-3a表达的变化。方法将35只健康成年SD大鼠用随机数字表法随机分为正常对照组5只和模型组30只，术前测量眼压。模型组大鼠一侧眼用Shareef—Sharma手术方法建立慢性高眼压模型，对侧眼只切开结膜，不烙闭巩膜表层静脉作为伪手术眼。按照术后不同处理时间随机将模型组分为术后1、3、5、7、14、28d组共6个亚组，每组5只。分别于术前及术后30min，1、3、7、14、28d用Tono—Pen接触式眼压笔测量双眼眼压。各模型组于术后各相应时间点与正常对照组分别取5只大鼠过量麻醉处死，制作视网膜石蜡切片行常规组织病理学检查，评价视网膜形态变化，用甲苯胺蓝染色法计数RGCs，采用免疫组织化学法检测各时间点各组大鼠Brn-3a在RGCs中的表达。结果高眼压模型组大鼠术后各时间点眼压均明显高于术前，差异有统计学意义（F=95．631，P=0．001），术后28d时眼压是正常对照组大鼠眼压的1．59倍。与伪手术眼相比，模型眼术后各时间点眼压均明显升高，差异均有统计学意义（q=18．418、15．261、10．987、6．931、4．975、2．962，P〈0．05）。正常对照组RGCs数量为（29．08±1．98）个／高倍视野，造模后3、5、7、14、28d组大鼠RGCs计数逐渐下降，差异均有统计学意义（t=5．943、8．034、15．023、17．004、19．371，P〈0．05）。免疫组织化学染色表明，随着造模时间的延长，各组Brn-3a阳性RGCs数量逐渐下降，差异有统计学意义（F=127．583，P=0．000）。结论采用Shareef—Sharma法可成功建立大鼠慢性高眼压动物模型，其眼压为中等程度升高；高眼压持续时�Background Brn-3a is a newly discovered specificity marker for retina ganglion cells（RGCs）. It is well-known that RGCs damage is a important pathological basis of hypertension-visual disorder. But the study concerning expression of Brn-3a in RGCs in glaucoma eye is still rately. Objective The purpose of this work was to investigate the changes of Brn-3a expression in model eye with chronic high intraocular pressure（IOP） and its relation with morphology of retina and the expression of Brn-3a in chronic ocular hypertension rats. Methods Thirty-five clean adult SD rats were randomly divided into normal control group （5 rats） and model group （30 rats）. Experimental chronic ocular hypertension models were induced unilaterally in the left eyes of 30 health adult SD rats by cauterizing super-scleral veins,and the conjunctival incision was made in the right eyes as sham operative group. The operated rats were subdivided into 6 groups according to the examination time points and 6 rats for each group.IOP was measured with Tono-Pen tonometer before and after 30 minutes, 1,3,7,14,28 day after surgery respectively. The rat models were sacrificed in 1,3,5,7,14,28 days after operation by excessive anesthesia method, and retinal section was prepared for the histopathological examination and the RGCs were counted using Nissl staining method. Expression of Brn-3a in RGCs was detected by immunohistochemistry. This experimental complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. Results The lOP was significantly raised from 30 minutes to 28 days after operation in model eyes with the top IOP（ 34.46±4.65 ）mmHg in the 30 minutes after operation, showing statistically significant differences in various time groups （ F = 95. 631, P = 0. 001 ） and different eyes （ F = 287. 473, P = 0. 001 ）. Compared with sham operative group, the IOP were elevated from 1 day through 14 days after operation （ q = 18. 418,15. 261,10. 987,

关 键 词：青光眼/模型 高眼压 视网膜神经节细胞 BRN-3A 

分 类 号：R774[医药卫生—眼科] R775[医药卫生—临床医学]