雷帕霉素对豚鼠形觉剥夺性近视的干预作用及相关机制研究  

Effects of rapamycin on the form-deprivation myopia in guinea pigs and its related mechanisms

作　　者：李欣蒙 高洪莲[2] 刘奇奇 于睿 张磊[1] LI Xinmeng;GAO Honglian;LIU Qiqi;YU Rui;ZHANG Lei(Department of Ophthalmology,the Affiliated Hospital of Binzhou Medical University,Binzhou 256600,Shandong Province,China;Clinical Medical Laboratory of Affiliated Hospital of Binzhou Medical University,Binzhou 256600,Shandong Province,China)

机构地区：[1]滨州医学院附属医院眼科,山东省滨州市256600 [2]滨州医学院附属医院临床医学实验室,山东省滨州市256600

出　　处：《眼科新进展》2021年第3期212-217,共6页Recent Advances in Ophthalmology

基　　金：山东省医药卫生科技发展计划项目(编号2016WS0022)。

摘　　要：目的探讨球结膜下注射雷帕霉素对豚鼠形觉剥夺性近视(FDM)的干预作用及相关机制。方法选取2~3周龄豚鼠80只(80眼),雌雄不限,随机分为4组空白对照组、FDM组、雷帕霉素组、FDM+雷帕霉素组;空白对照组不做任何处理;FDM组单纯缝合豚鼠右眼眼睑;雷帕霉素组于实验第1天、第7天豚鼠右眼球结膜下注射雷帕霉素50μg;FDM+雷帕霉素组于实验第1天、第7天豚鼠右眼球结膜下注射雷帕霉素50μg后再缝合右眼眼睑。记录各组豚鼠实验前及实验第7天、第14天的眼轴长度、屈光度;并取各组豚鼠视网膜行过碘酸-雪夫染色(PAS染色)及透射电镜检查,观察各组豚鼠视网膜形态变化;取各组豚鼠巩膜行RT-PCR检测巩膜组织中mTOR、基质金属蛋白酶-2(MMP-2)、α-平滑肌肌动蛋白(α-SMA)及转化生长因子β1(TGF-β1)的mRNA表达;利用Western blot检测各组豚鼠巩膜组织中AKT、p-AKT、mTOR、p-mTOR的蛋白表达。结果FDM组与空白对照组相比,实验第7天、第14天豚鼠的右眼眼轴长度、屈光度增加,形成了相对近视;FDM组巩膜中MMP-2、α-SMA的mRNA相对表达量与空白对照组相比均增加,TGF-β1 mRNA相对表达量减少,差异均具有统计学意义(均为P<0.05);与空白对照组相比,FDM组豚鼠巩膜中mTOR mRNA和AKT、P-AKT、mTOR、p-mTOR蛋白的相对表达量变化不大,差异均无统计学意义(均为P>0.05),但视网膜变薄,各层结构排列紊乱、疏松。雷帕霉素组与空白对照组相比,实验第7天、第14天豚鼠右眼眼轴长度、屈光度差异均无统计学意义(均为P>0.05),且视网膜形态无明显变化。FDM+雷帕霉素组与FDM组相比,实验第7天豚鼠右眼眼轴长度、屈光度差异均无统计学意义(均为P>0.05),实验第14天右眼眼轴长度和屈光度均减小,差异均有统计学意义(均为P<0.05),TGF-β1 mRNA的相对表达量增多,MMP-2、α-SMA、mTOR的mRNA和AKT、p-AKT、mTOR、p-mTOR的蛋白相对表达量均减少,差异均�Objective To investigate the effect of subconjunctival injection of rapamycin on the form-deprivation myopia(FDM)of guinea pigs and its related mechanisms.Methods Together 80 healthy guinea pigs(80 eyes),male or female,aged 2-3 weeks,were randomly divided into 4 groups blank control group,FDM group,rapamycin group,and FDM+rapamycin group.Guinea pigs in the blank control group left untreated;in the FDM group,the right eyelids of pigs were only sutured;1 day and 7 days of the experiment,50μg rapamycin was injected subconjunctival in the rapamycin group while the right eyelids were sutured after subconjunctival injection in the FDM+rapamycin group.The axial length and diopter of each group were recorded before and 7 days and 14 days of the experiment,and the eyeballs were taken for periodate Schiff’s staining(PAS staining)and retinal transmission electron microscope to observe the changes of retinal morphology.The sclera of each group of guinea pigs was taken for RT-PCR to measure mRNA expression of scleral tissue mTOR,matrix metalloproteinase-2(MMP-2),α-smooth muscle actin(α-SMA)and transforming growth factorβ1(TGF-β1);Western blot was used to detect the protein expression of AKT,p-AKT,mTOR,and p-mTOR in the scleral tissue of guinea pigs in each group.Results Compared with the blank control group,the right eyes of axial length and diopter in the FDM group increased on day 7 and 14 of the experiment,resulting in relative myopia.The relative expression of MMP-2 andα-SMA mRNA increased while the relative expression of TGF-β1mRNA decreased,and the differences were statistically significant(all P<0.05).Compared with the blank control group,there was no significant difference in the relative expression of mTOR mRNA and AKT,p-AKT,mTOR and p-mTOR protein in the sclera of guinea pigs in the FDM group(all P>0.05),and the retina became thinner and the structure of each layer becomes disordered and loose.There was no significant difference in axial length and diopter of right eye between rapamycin group and blank cont

关 键 词：形觉剥夺性近视 雷帕霉素 雷帕霉素靶蛋白 基质金属蛋白酶-2 Α-平滑肌肌动蛋白 转化生长因子β1 MTOR信号通路 

分 类 号：R778.1[医药卫生—眼科]