Long Evans大鼠视网膜缺血再灌注损伤模型的特征及其造模前后谷氨酸的含量  

Characteristics of the Long Evans model of retinal ischemia-reperfusion injury in rats and the content of glutamate before and after modeling

作　　者：熊亚妮 孟永 钱仪敏 张鹏 张慧 李华 XIONG Yani;MENG Yong;QIAN Yimin;ZHANG Peng;ZHANG Hui;LI Hua(Shanghai University of Engineering Science,Shanghai 201620,China;China State Institue of Pharmaceutical Industry,Shanghai InnoStar Biotech Co.,Ltd,Shanghai 201203;InnoStar Biotech Haimen Co.,Ltd,Haimen 226133;Shanghai Dingyue Biotechnology Co.,Ltd,Shanghai 201315)

机构地区：[1]上海工程技术大学,上海201620 [2]中国医药工业研究总院上海益诺思生物技术股份有限公司,上海201203 [3]益诺思生物技术南通有限公司,江苏海门226133 [4]上海鼎岳生物技术有限公司,上海201315

出　　处：《中国比较医学杂志》2023年第6期54-61,共8页Chinese Journal of Comparative Medicine

基　　金：江苏省创新能力建设计划科技公共服务平台项目(BM2021002)。

摘　　要：目的观察Long Evans大鼠视网膜缺血再灌注(retinal ischemia reperfusion,RIR)损伤后视网膜的功能、结构及谷氨酸含量的变化,为视网膜的损伤及可能保护机制研究提供参考。方法随机选取30只成年SPF级Long Evans大鼠,对其左眼前房持续60 min灌注高压生理盐水(132 mmHg),构建RIR损伤模型,右侧眼不处理作为自身对照。在造模后1、3、7和14 d,利用闪光全视网膜电图(flash electroretinogram,f-ERG)检测视网膜电生理功能的变化情况;在造模前及造模后3、7、14 d,利用光学相干断层技术(optical coherence tomography,OCT)测量视网膜厚度,眼底成像观察眼底血管的变化情况;于造模后14 d处死大鼠,进行石蜡包埋、苏木精伊红(hematoxylin eosin,HE)染色和缺口末端标记法(TdT-mediated dUTP nick end labeling,TUNEL)荧光染色观察视网膜形态结构、细胞凋亡及分布情况,ELISA检测视网膜谷氨酸的含量。结果与对照眼相比,造模眼从第1天开始视网膜电图b波振幅极显著下降(P<0.01),潜伏期极显著延迟(P<0.01);OCT显示从第3天开始视网膜神经节细胞复合体(retinal ganglion cell complex,GCC)厚度极显著变薄(P<0.01),从第7天开始全层视网膜厚度极显著变薄(P<0.01),且都随着时间延长越来越薄(P<0.05);眼底照片显示视网膜从第3天开始出现明显的缺血,一直到第14天都没有恢复到正常水平;第14天HE染色切片结果显示视网膜萎缩,内层明显变薄,视网膜神经节细胞(ganglion cells,RGCs)减少;TUNEL荧光染色结果显示视网膜各层出现明显的细胞凋亡;ELISA结果显示造模后视网膜谷氨酸含量增加(P<0.05)。结论Long Evans大鼠RIR损伤引起视觉电生理功能严重损伤,视网膜萎缩,尤其GCC厚度减少最明显,且随着时间延长损伤加剧,不可逆转,RGCs凋亡,眼底血管缺血,视网膜谷氨酸含量增加,为视网膜损伤类疾病的研究提供良好的动物模型。Objective To observe changes in retinal functions,structure,and glutamate content after retinal ischemia-reperfusion(RIR)injury in Long Evans rats to provide a reference for the study of retinal injury and possible protective mechanisms.Methods Thirty adult SPF level Long Evans rats were randomly selected,and the left anterior chamber of their left eye was perfused with high-pressure normal saline for 60 minutes to establish an RIR injury model,while the right eye was untreated as a control.At 1,3,7,and 14 days after modeling,changes in retinal electrophysiological functions were assessed by flash electroretinogram.The retinal thickness was measured by optical coherence tomography(OCT)before and at 3,7 and 14 days after modeling.Changes in fundus vessels were observed by fundus angiography.Rats were sacrificed at 14 days after modeling,and the retinal morphology,apoptosis,and distribution were observed by hematoxylin-eosin(HE)and TdT-mediated dUTP nick end labeling(TUNEL)staining.The content of glutamate in the retina was detected by ELISA.Results Compared with control eyes,the B wave amplitude of the electroretinogram in the model eyes were decreased significantly(P<0.01)and latency was delayed significantly(P<0.01)from the first day.OCT showed that the thickness of the retinal ganglion cell complex(GCC)was significantly thinner from day 3(P<0.01),the thickness of the whole retina was significantly thinner from day 7(P<0.01),and both of them became thinner over time(P<0.05).Fundus images showed that the retina had obvious ischemia from day 3 and did not recover to the normal level until day 14.On day 14,the HE staining showed retinal atrophy,obvious thinning of the inner layer and a reduction of retinal ganglion cells.TUNEL staining showed obvious apoptosis in all retinal layers.ELISAs showed that the glutamic acid content in the retina was increased after modeling(P<0.05).Conclusions RIR injury in Long Evans rat causes serious damage to visual electrophysiological functions,retinal atrophy,an obvious reduction

关 键 词：Long Evans大鼠 RIR损伤 视网膜 谷氨酸 动物模型 

分 类 号：R-33[医药卫生]