两种常用大鼠视神经钳夹伤模型造模效果的比较  

A comparative study of two common optic nerve crush models in rats

作　　者：安晶[1] 殷浩[1] 张作明[1] 

机构地区：[1]第四军医大学航空航天临床医学教研室,西安710032

出　　处：《中华实验眼科杂志》2016年第4期305-311,共7页Chinese Journal Of Experimental Ophthalmology

基　　金：陕西省科技资源开放共享平台项目（2015FWPT-02）;视觉系统疾病动物模型资源与功能鉴定平台

摘　　要：背景视神经钳夹伤（ONC）动物模型是外伤性视神经损伤致病机制及治疗方法相关基础研究的主要工具，目前常用的造模方法有经眼眶上缘开神经鞘膜视神经钳夹法和经球结膜外眦部夹伤视神经法，但关于2种模型优劣评价的研究很少。目的比较2种常用大鼠ONC模型的造模效果，为相关的实验研究中造模方法的选择提供依据。方法采用随机分配方法将8～10周龄健康成年雄性SD大鼠24只分为经眶上缘开神经鞘膜ONC组和经眶上缘开神经鞘膜ONC20s、40s、60s组，分别采用经眶上缘开神经鞘膜视神经夹伤法（20s）及经球结膜外眦部夹伤视神经法在大鼠的-侧眼建立ONC动物模型，各组大鼠的正常对侧眼作为对照。于造模后14d记录各组大鼠闪光视觉诱发电位（F—VEP）P。波；制备大鼠视神经组织切片，采用苏木精-伊红染色法观察大鼠视神经的组织病理学改变；采用免疫荧光法观察并计数大鼠视网膜组织中Brn-3a阳性视网膜神经节细胞（RGCs）。对2种造模方法的造模过程和检测结果进行比较。结果造模后14d，经眶上缘开神经鞘ONC组和经球结膜ONC20s组、40S组、60s组大鼠的F—VEPP。波潜伏期较各自的正常对侧眼均明显延长，差异均有统计学意义（t=-11．64、-8．04、-6．50、-10．84，均P〈0．01）；经眶上缘开神经鞘ONC组大鼠P．波潜伏期与经球结膜ONC20s、40s、60s组大鼠比较均明显延长，差异均有统计学意义（P=0．01、0．02、0．05）；各组大鼠术眼P．波振幅与其正常对侧眼比较差异均无统计学意义（均P〉0．05）。造模后14d，免疫荧光检测显示各组大鼠模型眼视网膜上Brn-3α表达阳性RGCs数均较正常对侧眼明显减少，经眶上缘开神经鞘ONC组大鼠视网膜上Brn-3a表达阳性RGCs数为（13．60±2．14）个／视野，为其正常对侧眼的47．49％，经球结膜ONC20s、40s和60s组中Brn-3a阳性RGCs数�Background Optic nerve crush （ONC） model is an available tool in the basic research on the mechanism and treatment of optic nerve injury. The opening optic nerves sheath crushing and via bulbar conjunctiva lateral canthus optic nerves crushing are frequently used ONC modeling methods. However, the comparison between these models is not elaborated. Objective This study was to compare the outcomes between opening sheath ONC model and via bulbar conjunctiva lateral canthus ONC model. Methods Twenty-four male SD rats were randomized into four groups. ONC models were established via superorbital rim opening sheath to crushing optic nerve for 20 seconds in the opening sheath ONC group,or via bulbar conjunctiva lateral canthus cutting to crush optic nerve for different time in the via bulbar conjunctiva ONC 20-second, 40-second and 60-second groups, respectively. All models were monocular created in the rats, and the fellow eyes served as controls. In 14 days after modeling,flash visual evoked potential （F-VEP） were recorded, optic nerve and retinal sections were prepared in the rats. The histopathology of the samples was examined by hematoxylin and eosin staining. The expression of Brn-3a in the retinalganglion cells （RGCs） was detected by immunofluorescence technique and the number of Brn-3a RGCs was counted. The modeling procedure and outcomes were compared between the two approaches. Results The latencies of P waves were significantly extended in the opening sheath ONC group, via bulbar conjunctiva ONC 20-second, 40-second and 60-second groups in comparison with the corresponding control eyes （t =- 11.64,-8.04,-6.50, -10.84 ,all at P〈0.01 ）. The P1 latencies were longer in the opening sheath ONC group than those in the via bulbar conjunctiva ONC 20-second,40-second and 60-second groups （ P= 0. 01,0. 02,0.05 ） , but no significant differences were found in the amplitudes of Pl waves between model eyes and corresponding control eyes （ all at P〉0.05）. The Brn-3a RGCs numbers were evidently d

关 键 词：视神经/损伤 视神经/病理学 闪光视觉诱发电位 视网膜神经节细胞/病理 动物模型 

分 类 号：R779.1[医药卫生—眼科]