细胞周期依赖性蛋白激酶5抑制剂Roscovitine对皇家外科学院大鼠视网膜光感受器细胞凋亡的影响  

Effects of Cdk5 inhibitor Roscovitine on retinal degeneration in Royal College of Surgeons rat

作　　者：任英华[1] 邹刚[1] 张金金[1] 盛迅伦[1] 

机构地区：[1]宁夏回族自治区人民医院眼科宁夏眼科医院,银川750001

出　　处：《中华眼底病杂志》2015年第3期263-267,共5页Chinese Journal of Ocular Fundus Diseases

基　　金：国家自然科学基金（81060078）

摘　　要：目的 观察细胞周期依赖性蛋白激酶5（Cdk5）抑制剂Roscovtine对皇家外科学院（RCS）大鼠视网膜光感受器细胞凋亡的影响.方法 RCS雄性大鼠30只,随机数字表法随机分为早期干预组、中期干预组、晚期干预组,每组10只.分别于出生后17、25、35 d,给予右眼玻璃体腔注射Cdk5抑制剂Roscovitine 4μl作为实验眼;左眼玻璃体腔注射Roscovitine溶剂二甲基亚砜4μl作为对照眼.早期干预组、中期干预组、晚期干预组大鼠分别于出生后25、35、45 d处死,摘除眼球作视网膜切片,采用苏木精-伊红（HE）染色观察视网膜外核层厚度变化;采用免疫组织化学染色观察Cdk5、p25及活化型半胱氨酸天冬氨酸蛋白酶3 （cleave-caspase 3）的阳性表达;采用原位末端标记法（TUNEL）检测视网膜凋亡细胞的表达.取视网膜组织,采用蛋白免疫印迹法（Western blot）检测大鼠视网膜中Cdk5、p25及cleave-caspase 3的蛋白表达.对Cdk5、p25、cleave-caspase 3阳性表达与凋亡细胞表达两两变量之间行多变量偏相关分析.结果 早期干预组、中期干预组大鼠实验眼外核层厚度均较对照眼厚,且早期干预组实验眼与对照眼外核层 厚度的差异更明显.晚期干预组大鼠实验眼与对照眼的外核层厚度差异不明显.免疫组织化学染色结果显示,早期干预组、中期干预组实验眼外核层Cdk5、p25、cleave-caspase 3阳性表达均较对照眼降低,差异均有统计学意义（P＜0.05）.晚期干预组实验眼与对照眼外核层Cdk5、p25、cleave-caspase 3阳性表达比较,差异无统计学意义（P＞0.05）.TUNEL检测结果显示,早期干预组、中期干预组大鼠实验眼视网膜外核层凋亡细胞表达较对照眼明显降低;晚期干预组大鼠实验眼与对照眼视网膜外核层凋亡细胞表达差异不明显.多变量偏相关分析结果显示,Cdk5、p25、cleave-caspase 3阳性表达与凋亡细胞表达两两变量之间均呈明显相关性（P＜0Objective To evaluate the photoreceptor-protective effects of Cdk5 inhibitor Roscovitine on retinal degeneration in Royal College of Surgeons （RCS） rat.Methods The RCS rats were divided into three groups according to postnatal days：the early （17 days）,medium （25 days） and late intervention group （35 days）.Cdk5 inhibitor Roscovitine were used in the right eyes by intravitreal injection as experimental eyes and Roscovitine solvent dimethylsulfoxide were used in the left as control at postnatal 17,25,35 days.Hematoxylin-eosin （HE） staining was used to observe the thickness of outer nuclear layer.The expression of Cdk5 P25 and cleave-caspase 3 in the retina was evaluated by immunohistochemistry.The protein expression of cleave-caspase 3 in the retina was determined by Western blot.The apoptosis of retinal cells was examined by terminal deoxynucleotidyl transferase mediated nick end labeling.Results HE staining showed that thickness of outer nuclear layer in the early and medium intervention groups were significantly thicker than that in the control group （P〈0.05）,particularly in the early intervention group.And there was no significant change in the late intervention group （P〉0.05）.The expression level of Cdk5,p25,cleave-caspase 3 in the outer nuclear layer in three intervention groups were lower than that in the control group （P〈0.05）,especially in the early intervention group.Conclusion Cdk5 inhibitor Roscovitine can delay the retinitis pigmentosa process in RCS rats by early,medium interventional therapy and may have a certain degree of photoreceptor-protective effects.

关 键 词：细胞周期蛋白依赖激酶5/拮抗剂和抑制剂 光感受器细胞/病理学 细胞凋亡 动物实验 

分 类 号：R774.1[医药卫生—眼科]