核黄素-紫外线A巩膜胶原交联照射时间与视网膜损伤的关系  

Relationship between irradiation time of riboflavin-ultraviolet A scleral collagen cross-linking and retinal injury

作　　者：颜东 刘丽梅 明春秀 张少斌 Yan Dong;Liu Limei;Ming Chunxiu;Zhang Shaobin(Binzhou Medical University,Yantai 264033,China;Weifang Eye Hospital,Weifang 261041,China)

机构地区：[1]滨州医学院,烟台264033 [2]潍坊眼科医院,261041

出　　处：《中华实验眼科杂志》2021年第6期483-489,共7页Chinese Journal Of Experimental Ophthalmology

基　　金：潍坊市科技发展计划项目(2017YX031)。

摘　　要：目的研究不同照射时间下核黄素-紫外线A巩膜胶原交联术对兔视网膜的影响,寻找照射的安全时间。方法采用随机数表法将60只健康成年新西兰大白兔随机分为对照组(0 min组)和照射10、20、30及40 min组,每组12只,均取左眼进行核黄素-紫外线A巩膜胶原交联照射(370 nm,10 mW/cm 2)。采用光学显微镜和透射电子显微镜观察并比较各组视网膜病理组织学改变;采用生化反应试剂盒检测并比较各组视网膜组织中丙二醛(MDA)浓度及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)活性;采用Western blot法检测并比较各组视网膜组织中SOD、CAT蛋白表达水平。结果光学显微镜下观察到对照组视网膜各层结构排列有序,照射10 min组与对照组相比无明显变化,随着照射时间的延长,视网膜结构损伤逐渐加重。透射电子显微镜下可见对照组光感受器细胞外节膜盘层状结构完整,光感受器细胞内节线粒体结构完整,线粒体嵴结构连续,照射10 min组与对照组相比无明显变化,随着照射时间的延长,视网膜结构损伤逐渐加重。各组视网膜组织中MDA浓度随着照射时间的延长逐渐升高,总体比较差异有统计学意义(F=65.25,P<0.05),其中照射20 min、30 min、40 min组MDA浓度分别为(11.31±1.84)、(14.94±1.04)和(18.25±1.42)nmol/mgprot,明显高于对照组的(1.13±0.02)nmol/mgprot,差异均有统计学意义(均P<0.05);照射20 min、30 min、40 min组MDA浓度依次升高,两两比较差异均有统计学意义(均P<0.05)。随着照射时间的延长,视网膜组织中SOD、CAT和GSH-Px活性及SOD、CAT蛋白相对表达量均逐渐降低,总体比较差异均有统计学意义(F=44.09、34.18、35.60、115.75、78.86,均P<0.05),其中照射20、30、40 min组与对照组比较,照射20 min、30 min、40 min组两两比较,差异均有统计学意义(均P<0.05)。结论10 mW/cm 2核黄素-紫外线A巩膜胶原交联照射兔眼10 min是安�Objective To investigate the effects of riboflavin-ultraviolet A scleral collagen cross-linking on the retina under different irradiation time,and to determine the safe irradiation time.Methods Sixty healthy New Zealand white rabbits were randomly divided into control group(0 minute group),10 minutes group,20 minutes group,30 minutes group and 40 minutes irradiation group according to the irradiation time,with 12 rabbits in each group.The left eye was irradiated with riboflavin-ultraviolet A scleral collagen(370 nm,10 mW/cm 2).The histopathological change of retina was observed by light microscope and transmission electron microscope and compared among different groups.The concentration of MDA and the activities of SOD,CAT and GSH-Px in retinal tissue were detected by corresponding kits.The expression levels of SOD and CAT proteins in retinal tissue were detected by Western blot method.The study protocol was approved by the Binzhou Medical University Laboratory Animal Ethical Committee(No.2017-80).The use and care of animals complied with the statement of ARVO and the Regulation on the Management of Laboratory Animal Quality of China.Results Under the light microscope,the structure of the retinas in the control group was orderly arranged.Under the transmission electron microscope,the lamellar structure in the inner segment and the mitochondrial structure in the outer segment of the photoreceptor cells were intact,and the mitochondrial ridge was continuous in the control group.There was no obvious difference in retinal morphology between the 10 minutes irradiation group and the control group under both the light microscope and the transmission electron microscope,and the retinal damage became more severe with the prolongation of irradiation time.The concentration of MDA in the retina of each group was elevated gradually with the increase of irradiation time,and the difference was statistically significant(F=65.25,P<0.05).The concentration of MDA was(11.31±1.84),(14.94±1.04)and(18.25±1.42)nmol/mgprot in the 20 m

关 键 词：胶原交联 巩膜/药物作用 核黄素/药理 紫外线/不良反应 光感受器细胞/药物作用 光敏剂/毒性作用 实验性放射损伤/病理 超微结构 脂质过氧化损伤 

分 类 号：R774.1[医药卫生—眼科]