背景 以往国内关于青光眼研究的造模方法中大多存在高眼压维持时间较短的问题.目前前房内注射聚苯乙烯微球建立小鼠慢性高眼压模型的方法已在国外应用,但国内尚缺乏对该模型的评价.目的 评价前房内一次性注射聚苯乙烯微球建立小鼠慢性高眼压模型的方法学及应用价值.方法 将42只SPF级成年C57BL/6L雌性小鼠用随机数字表法随机分为3个组,正常对照组小鼠不进行任何处理;PBS组小鼠前房内一次性注射PBS溶液2μl;微球组小鼠左眼前房内一次性注射聚苯乙烯微球2μl.各组小鼠均于注射后2周和4周行裂隙灯显微镜检查,采用TonoLab回弹式眼压计测量眼压;并于相应时间点观察结束后处死动物,制备眼球的组织学切片和视网膜铺片,在光学显微镜下观察前房角情况;采用质量分数4%荧光金经上丘逆行标记视网膜神经节细胞(RGCs),在荧光显微镜下计数各组小鼠视网膜铺片中RGCs的存活密度和神经纤维的数量;采用免疫组织化学染色法检查视网膜铺片中β-Ⅲ-微管蛋白阳性细胞数. 结果 前房注射后2周和4周,微球组小鼠眼压均明显高于正常对照组和PBS组,差异均有统计学意义(P<0.05).微球注射后2周小鼠平均眼压达高峰,为(29.67±2.34)mmHg(1 mmHg=0.133 kPa),之后眼压逐渐下降,4周时小鼠的平均眼压为(15.71±1.23)mmHg,但明显高于术前,差异均有统计学意义(P<0.05).微球组小鼠前房注射后裂隙灯显微镜下可见角膜水肿,但前房内未见明显的炎症反应,眼球组织病理学检查可见微球聚集在前房角.前房注射后2周和4周,微球组RGCs存活密度分别为(4 542.82±653.72)个/mm2和(3 623.12±628.79)个/mm2,明显低于正常对照组的(6 979.33±678.49)个/mm2和(6 963.91±497.29)个/mm2,差异均有统计学意义(t=17.729、28.569,P<0.05),亦明显低于PBS组的(6 843.21 ±573.42)个/mm2和(6 937.53±465.24)个/mm2,?
Background Many methods of ocular hypertension modeling have been used before,but these models remain short-duration ocular hypertension only.A new method of elevating intraocular pressure in mice by anterior chamber injection of polystyrene microspheres was reported abroad.However,this model is rarely used in China.Objective This study was to evaluate the application value of anterior chamber injection of polystyrene microspheres to establish glaucoma model in mice.Methods Forty-two SPF adult female C57BL/6L mice were divided into three groups according to random number table.Polystyrene microspheres (2 μl) were injected into the anterior chamber monocularly in the microspheres group,and the equal amount of PBS was used in the same way in the PBS group.No intervene was performed in the normal control group.The eyes of mice were examined by slit lamp microscope,and the intraocular pressure (IOP) was measured with TonoLab rebound tomometer in a 3-day interval after injection.Ocular histological sections were prepared 2 and 4 weeks after injection,and the anterior chamber angle was examined under the optical microscope.Neurons retrograde labeling was performed by 4% fluorogold to calculate the survival number of retinal ganglion cells (RGCs) and the nerve fiber density was detected to assess the degree of RGCs and axon damage in retinal flat mounts,and the β-Ⅲ-tubulin-positive cells in the RGCs layer were examined by immnofluorescence method.The use and care of the animals complied with the instruction of Association for Research in Vision Ophthalmology (ARVO).Results IOP was significantly higher in the mice of the microspheres group than that in the normal control group or PBS group 2 and 4 weeks (all at P〈0.05).In the microspheres group,IOP reached peak in 2 weeks after injection and was significantly higher than that of 4 weeks after injection ([29.67±2.34] mmHg versus[15.71±1.23] mmHg) (all at P〈0.05).In 2 and 4 weeks after the anterior chamber injection of polystyrene,corn