聚苯乙烯微球前房注射诱导小鼠高眼压模型  

Elevated intraocular pressure mice model induced by anterior chamber injection of polystyrene microspheres

作　　者：魏欣[1] 陈晓明[1] 佘春燕[1] 李妮[1] 霍朝奎[1] 刘旭阳[2] 

机构地区：[1]四川大学华西医院眼科,四川省成都市610041 [2]暨南大学医学院深圳眼科中心,广东省深圳市510632

出　　处：《眼科新进展》2013年第3期204-209,共6页Recent Advances in Ophthalmology

基　　金：国家自然科学基金青年基金(编号:81200687);教育部博士点基金(编号20120181120014);四川省成都市科技支撑项目(高校院所应用成果转化)(编号:12DXYB058JH-002)~~

摘　　要：目的建立一种新型的青光眼小鼠模型,为青光眼发病机理和治疗的研究打下基础。方法成年C57BL/6J小鼠107只随机分为5组:A组接受15μm聚苯乙烯微球前房注射(n=28);B组接受10μm微球注射(n=44);C组分别于0d及23d接受2次10μm微球注射(n=15);D组为对照组,接受PBS前房注射(n=10);E组为空白对照(n=10);所有鼠右眼注射。注射后每2d用TonoLab眼压计测量眼压1次以监测眼压变化。在眼压持续升高14d、28d或56d时,采用电镜、荧光金逆行标染以及免疫组织化学的方法对视网膜神经节细胞和轴突变性的情况进行定量评估并比较;同时在眼压升高14d、28d时,用双重标染的方法对比荧光金及β-Ⅲ-tubulin免疫标染视网膜神经节细胞的结果有无差异。结果监测双眼眼压,D组小鼠在实验期间眼压值稳定,为(10.3±1.6)mmHg(1kPa=7.5mmHg),与E组(10.0±1.5)mmHg相比差异无统计学意义;87只接受微球前房注射的小鼠眼有81只较对照眼升高。A组注射15μm微球后可以诱导眼压上升约14d,峰值为(21.8±2.1)mmHg。单次注射10μm直径微球可以诱导眼压上升28d,峰值达到(24.4±6.2)mmHg;在23d时进行第2次注射,高眼压状态可以维持到56d。B组14d、28d及C组首次注射后56d,与对侧眼视神经截面相比,轴突丢失率分别为(22.4±2.1)%、(28.0±3.1)%和(42.0±3.8)%;而同样时间点下视网膜神经节细胞的丢失率分别为(23.8±2.3)%、(35.8±4.4)%及(45.3±3.9)%,各组间差异均有统计学意义。双重标染β-Ⅲ-tubulin和荧光金发现,β-Ⅲ-tubulin阳性细胞的数量分别为(3150±30)个·mm-2(14d)和(2574±165)个·mm-2(28d),而荧光金阳性细胞数量分别(3040±465)个·mm-2(14d)和(2433±192)个·mm-2(28d),各组间差异均无统计学意义。结论前房注射微球能够高效且稳定地诱导小鼠眼压升高和青光眼视神经病变,是一种较理想的新型青光眼模型。Objective To establish a new glaucoma mice model for studying the pathogenesis and treatment of glaucoma. Methods A total of 107 adult C57BL/SJ mice were divided into 5 groups randomly ：group 1 accepted 15 μm polystyrene micro- spheres anterior chamber injection（ n = 28 ）, group 2 accepted 10 μm microspheres ante- rior chamber injection（ n -- 44）, group 3 accepted 10 μm microspheres anterior chamber twice injection at 0 day and 23 days（n = 15 ）, group 4 was control group, accepted PBS anterior chamber injection（ n = 10）, group 5 was blank control group （ n = 10 ）, the right eye in all mice received injection. Intraocular pressure （IOP） was measured every other day using a TonoLab tonometer. After maintaining an elevated IOP for 14 days,28 days or 56 days, the degree of retinal ganglion cells （RGC） and axon degeneration was as- sessed quantitatively by electron microscope, FluoroGold retrograde labeling and immu- nohistochemistry. Meanwhile, double labeling was used to compare whether any signifi- cant difference between FluoroGold labeling and β-Ⅲ-tubulin immunolfistochemistry la- beling after high IOP at 14 days and 28 days. Results IOP in group 4 kept stable, which was （ 10.3 ± 1.6） mmHg（ 1 kPa =7.5 mmHg） ,there was no significant difference compared with group 5 （ 10.0 ±1. 5） mmHg;S1 out of 87 mice that received anterior chamber injection of microspheres exhibited consistent IOP elevation above that of con- trol eyes. Injection of 15 μm microspheres resulted in a 14 days elevation of IOP,which peak IOP value was（21.8 ± 2.1 ）mmHg; 10 μm microspheres injection resulted in a 28 days elevation of IOP, which peak IOP value was （24.4 ± 6.2） mmHg. High IOP could be maintained to a 56 days period after a second injection of microspheres in week 4. As the duration of IOP elevation increased, both RGC bodies and their axons degenerated progressively. After high IOP last forl4 days,28 days and 56 days,the loss rate of axon density were （ 22.4±2.1 ） % , �

关 键 词：青光眼 聚苯乙烯微球 视网膜神经节细胞 轴突变性 高眼压模型 鼠 

分 类 号：R988.1[医药卫生—药品]