视黄醇脱氢酶5在豚鼠形觉剥夺性近视眼中的表达变化  

Differences of retinol dehydrogenase 5 expression in form-deprivation myopic eyes of guinea pigs

作　　者：毛玉梅 兰长骏[1] 杨琴 钟维琪 谷志明 周桂梅 廖萱[1] MAO Yumei;LAN Changjun;YANG Qin;ZHONG Weiqi;GU Zhiming;ZHOU Guimei;LIAO Xuan(Department of Ophthalmology,Affiliated Hospital of North Sichuan Medical College,Department of Ophthalmology&Optometry,North Sichuan Medical College,Nanchong 637000,Sichuan Province,China)

机构地区：[1]川北医学院附属医院眼科,川北医学院眼视光学系,四川省南充市637000

出　　处：《眼科新进展》2021年第11期1011-1015,共5页Recent Advances in Ophthalmology

基　　金：四川省科技计划项目(编号2019YJ0381);四川省医学会科研课题(编号S20016)。

摘　　要：目的探讨形觉剥夺性近视(FDM)和正常豚鼠的视网膜色素上皮(RPE)细胞中的视黄醇脱氢酶5(RDH5)基因在转录水平和蛋白水平的表达差异。方法30只豚鼠随机分为实验组(15只),其中,右眼进行遮盖为FDM组(15眼),左眼不遮盖为自身对照组(15眼);空白对照组(15只)双眼不做任何干预。分别于遮盖前,遮盖1周、2周、3周和4周后检查豚鼠屈光度和眼轴长度。采用免疫荧光染色法检测各组豚鼠RPE细胞中RDH5表达,实时荧光定量PCR和Western blot检测各组豚鼠RPE细胞中RDH5 mRNA和蛋白的表达变化。结果遮盖前、遮盖1周后,FDM组、自身对照组、空白对照组三组间豚鼠的屈光度、眼轴长度差异均无统计学意义(均为P>0.05);遮盖2周、3周、4周后,三组间豚鼠的屈光度、眼轴长度差异均有统计学意义(均为P<0.05)。遮盖2周、3周、4周后,FDM组豚鼠屈光度分别为(1.47±1.41)D、(3.32±1.39)D、(4.63±1.04)D,眼轴长度分别为(7.69±0.24)mm、(7.92±0.09)mm、(8.34±0.15)mm,与空白对照组和自身对照组相比,差异均有统计学意义(均为P<0.05)。遮盖4周后,与空白对照组和自身对照组相比,FDM组豚鼠RPE细胞中RDH5 mRNA水平和蛋白水平均明显下调(均为P<0.05)。免疫荧光染色结果显示,RDH5主要表达于豚鼠RPE层,与空白对照组和自身对照组相比,FDM组豚鼠RPE层RDH5蛋白的平均荧光强度最低,差异均有统计学意义(均为P<0.05)。结论FDM豚鼠近视眼RPE细胞内RDH5基因在转录和蛋白水平的表达均下调,提示RPE细胞内RDH5可能在近视的发生发展中扮演了一定的角色。Objective To explore the difference of the retinol dehydrogenase 5(RDH5)gene expression at the transcriptional and protein levels in the retinal pigment epithelium(RPE)between form-deprivation myopic(FDM)guinea pigs and normal guinea pigs.Methods Thirty guinea pigs were randomly assigned to two groups the experimental group(n=15),with 15 right eyes covered(FDM group)and 15 left eyes not covered(self-control group);and the blank control group(n=15),with all eyes left uncovered.Diopter and axial length were examined before covering,1,2,3,and 4 weeks after covering,respectively.The immunofluorescence staining was used to detect the expression of RDH5 in each group.The quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot were used to detect the changes in RDH5 mRNA and protein expression.Results There were no statistically significant differences in diopter and axial length among the three groups before eyes were covered and one week after eyes were covered(all P>0.05).However,after two,three and four weeks of covering,the differences in diopter and axial length of the three groups showed statistical significance(all P<0.05).After two,three and four weeks of covering,the diopter of guinea pigs in the FDM group was(1.47±1.41)D,(3.32±1.39)D,and(4.63±1.04)D,respectively,and the axial length was(7.69±0.24)mm,(7.92±0.09)mm,and(8.34±0.15)mm,respectively,showing statistically significant differences compared with the blank control group and self-control group(all P<0.05).After four weeks,compared with the blank control group and the self-control group,the RDH5 mRNA and protein levels in the FDM group were significantly downregulated(all P<0.05).Immunofluorescence staining showed that RDH5 was mainly expressed in the RPE layer.The FDM group had the lowest average fluorescence intensity of RDH5 expression in the RPE layer,and the differences were statistically significant(all P<0.05).Conclusion Both transcriptional and protein expression of RDH5 in RPE cells of the FDM guinea pigs are downregulated,

关 键 词：豚鼠 近视 形觉剥夺 视网膜色素上皮 视黄醇脱氢酶5 

分 类 号：R778[医药卫生—眼科]