横向定量牵拉法制作大鼠视神经精确损伤模型

2016-09-02谷新怡赵朋波刘爱伟尚姗姗闫晓玲吴鲁华夏燕婷

国际眼科杂志 2016年9期

谷新怡，周　剑，赵朋波，刘爱伟，尚姗姗，闫晓玲，吴鲁华，夏燕婷

·实验论著·

横向定量牵拉法制作大鼠视神经精确损伤模型

谷新怡1，周剑1，赵朋波1，刘爱伟1，尚姗姗2，闫晓玲1，吴鲁华2，夏燕婷1

Foundation items:National Natural Science Foundation Project (No.81173307); Natural Science Foundation Project of Beijing (No.16G40086)

1Department of Ophthalmology, Dongfang Hospital Affiliated to Beijing University of Chinese Medicine, Beijing 100078, China;2Department of Ophthalmology, the Third Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100029, China

Correspondence to:Jian Zhou. Department of Ophthalmology, Dongfang Hospital Affiliated to Beijing University of Chinese Medicine, Beijing 100078, China.zhj9667@126.com

Abstract

•AIM: To make a rat model of optic nerve injury by using the transverse quantitative traction method, and to evaluate the survival rate of retinal ganglion cells (RGCs) of the model using the method of fluorogold retrograde labeling.

•METHODS:Twenty-five Wistar rats were randomly divided into five groups: sham operation group, 1d, 3d, 7d and 14d after tractive optic nerve injury group. The model groups pulled the left optic nerve with lateral tensiometer; the sham operation group only exposed the optic nerve but not pulled. The right eyes of each group were served as normal control eyes. The RGCs density of the five groups was observed by fluorogold retrograde labeling.

•RESULTS:In the normal control group, the RGCs labeled by the fluorescent gold were round or oval, clear boundary, no obvious fluorescent dye leakage and partially visible cell processes. However, in the optic nerve traction groups, the number of RGCs decreased with time increasing and the cell distribution was not uniform. Lots of fluorescent leakages and microglial cells were observed. Compared with the normal control group, there was no significant difference in the morphology and density of RGCs of sham operation group (P>0.05). In 1d, 3d, 7d and 14d after traction of the optic nerve groups, the number of RGCs were reduced progressively and the density of RGCs of the left eye was significantly lower than that in the normal control group (P<0.01). The survival rates of RGCs in the groups of 1d, 3d, 7d and 14d after optic nerve traction were (78.94±0.92)%, (60.07±0.90)%, (38.92±1.42)% and (17.31±0.97)% respectively.

•CONCLUSION:The transverse quantitative traction method can establish a model of easily quantifiable optic nerve injury, which can provide a powerful tool for further study on the treatment of optic nerve injury.

目的：采用横向定量牵拉法制作大鼠视神经损伤模型，并利用荧光金逆行标记评价视神经牵拉伤后视网膜节细胞(retinal ganglion cells，RGCs)的存活率。

方法：将25只雄性Wistar大鼠随机均分为5组，即假手术组和视神经牵拉伤后1、3、7、14d组。模型组用横向张力计牵拉左眼视神经，假手术组仅暴露左眼视神经但不予牵拉，各组以右眼为正常对照。用荧光金逆行标记，并观察假手术组及视神经牵拉伤后1、3、7、14d组RGCs的密度。

结果：正常对照组RGCs形态多呈圆形或椭圆形，边界清楚，细胞外无明显荧光染料渗漏，部分可见明显细胞突起；而视神经牵拉伤后RGCs随时间延长而不断减少，细胞分布不均匀，并可见大量荧光渗漏及小胶质细胞。与正常对照组相比，假手术组RGCs形态和密度无明显差异(P>0.05)；视神经牵拉伤后第1、3、7、14d的RGCs数量进行性减少，且其密度均明显低于正常对照组(P<0.01)；视神经牵拉伤后第1、3、7、14d的RGCs存活率分别为78.94%±0.92%、60.07%±0.90%、38.92%±1.42%和17.31%±0.97%。

结论：横向定量牵拉法可以建立易于量化的视神经损伤模型，为进一步研究视神经损伤后的治疗方法提供有力工具。

横向定量牵拉法；视神经损伤；荧光金；视网膜神经节细胞

引用：谷新怡，周剑，赵朋波，等.横向定量牵拉法制作大鼠视神经精确损伤模型.国际眼科杂志2016;16(9):1625-1628