Krüppel样因子7对视网膜缺血再灌注损伤小鼠视网膜神经节细胞存活及视网膜电图的影响  

Effects of Krüppel-like factor 7 on the survival of retinal ganglion cells and electroretinogram after retinal ischemia-reperfusion injury

作　　者：李宗媛 杨宁[1] 罗晋媛 杨家翼 贺涛[1] 邢怡桥[1] Li Zongyuan;Yang Ning;Luo Jinyuan;Yang Jiayi;He Tao;Xing Yiqiao(Eye Center,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院眼科中心,430060

出　　处：《中华眼底病杂志》2020年第11期846-852,共7页Chinese Journal of Ocular Fundus Diseases

基　　金：国家自然科学基金面上项目(81271025);中央高校基本科研业务费专项资金资助(青年教师资助项目)(2042020kf0065);湖北省自然科学基金面上项目(青年)(2020CFB240)。

摘　　要：目的观察Krüppel样因子7(KLF7)对视网膜缺血再灌注(RIR)损伤小鼠RGC存活及ERG的影响。方法采用随机数字表法将126只雄性C57BL/6J小鼠随机分为正常组、RIR组、正常-KLF7组、正常-绿色荧光蛋白(GFP)组、RIR-KLF7组、RIR-GFP组。小鼠8周龄时,正常-KLF7组及RIR-KLF7组小鼠玻璃体腔注射1.0×1012 vg/ml过表达KLF7的重组腺相关病毒载体(AAV2-KLF7-GFP)1μl;正常-GFP组及RIR-GFP组小鼠玻璃体腔注射同样滴度的仅带GFP的空白腺相关病毒载体1μl。小鼠11周龄时,RIR组、RIR-KLF7组、RIR-GFP组小鼠建立RIR模型并测量眼压。玻璃体腔注射AAV2-KLF7-GFP病毒载体4周后,利用视网膜冰冻切片观察病毒转染情况。RIR建模后第7天,利用视网膜铺片免疫荧光染色定量观察RGC存活率;行全视野ERG检查,观察其暗适应a、b波和振荡电位(Ops)、光负性反应(PhNR)振幅和潜伏期的变化;行视动反应检测,观察其视敏度的变化。玻璃体腔注射病毒4周后,采用Western bolt检测小鼠视网膜中KLF7的蛋白表达。结果与正常组比较,RIR组小鼠视网膜RGC存活率,ERG a、b波振幅,Ops、PhNR振幅以及视敏度均明显降低,差异有统计学意义(t=12.860、7.157、5.735、8.953、4.744、9.887,P<0.05)。随着刺激光强的增加,小鼠暗适应ERG a、b波振幅逐渐升高,其潜伏期逐渐缩短。与RIR组比较,RIR-KLF7组小鼠视网膜RGC存活率,ERG a、b波振幅,Ops、PhNR振幅以及视敏度均明显升高,差异有统计学意义(t=6.350、3.253、3.695、5.825、5.325、4.591,P<0.05)。Western bolt检测结果显示,正常-KLF7组小鼠视网膜中KLF7蛋白表达较正常组明显上调,差异有统计学意义(t=4.105,P<0.01)。结论KLF7过表达可提高RGC存活率,保护其电生理功能。Objective To investigate the effects of Krüppel-like factor 7 (KLF7) on the survival of retinal ganglion cells (RGCs) and electroretinogram (ERG) after retinal ischemia-reperfusion (RIR) injury in mice.Methods A total of 126 male C57BL/6J mice were randomly divided into normal group, RIR group, normal-KLF7 group, normal-green fluorescent protein (GFP) group, RIR-KLF7 group and RIR-GFP group. At the age of 8 weeks, mice of normal-KLF7 group and RIR-KLF7 group were intravitreally injected 1ul of 1.0×10^12 vg/ml adeno-associated virus overexpressing KLF7 (AAV2-KLF7-GFP). Mice of normal-GFP group and RIR-GFP group were injected adeno-associated virus of AAV2-GFP with the same titer. At the age of 11 weeks, RIR injury was induced in mice of RIR group, RIR-KLF7 group and RIR-GFP group, and intraocular pressure was measured. Retinal cryosections were used to access the efficacy of virus transfection 4 weeks after AAV2-KLF7-GFP transfer. 7 days after RIR injury, RGCs' survival rate was observed and quantified by immunofluorescent staining. ERG was performed to observe the differences in amplitudes and incubation period of scotopic ERG a-, b-wave, oscillatory potentials (Ops), photopic negative responses (PhNR). Optomotor response was performed to observe the differences of visual acuity. Expression of KLF7 was detected by western blot 4 weeks after AAV2-KLF7-GFP transfer.Results Compared with normal group, RGCs’ survival rates, amplitudes of ERG a-, b-wave, Ops, PhNR and visual acuity of mice in RIR group were decreased, and the differences were statistically significant (t=12.860, 7.157, 5.735, 8.953, 4.744, 9.887;P<0.05). With the increase of light intensity, the amplitudes of scotopic ERG a- and b-wave were gradually increased while the incubation period was gradually shortened. Compared with RIR group, RGCs’ survival rates, amplitudes of ERG a-, b-wave, Ops, PhNR and visual acuity of mice in RIR-KLF7 group were increased, and the differences were statistically significant (t=6.350, 3.253, 3.695, 5.825, 5.325, 4

关 键 词：Kruppel样转录因子类 再灌注损伤 视网膜神经节细胞 视网膜电描记术 动物实验 

分 类 号：R774.1[医药卫生—眼科]