急性高眼压诱导的小鼠血-视网膜外屏障破坏  

Breakdown of blood-retinal outer barrier in mice following acute high intraocular pressure

作　　者：李雯 曹兴[1] 叶长华 Li Wen;Cao Xing;Ye Changhua(Aier School of Ophthalmology,Central South University,Changsha Aier Eye Hospital,Changsha 410004,China;Department of Ophthalmology,Hunan Children's Hospital,Changsha 410007,China)

机构地区：[1]中南大学爱尔眼科学院长沙爱尔眼科医院,长沙410004 [2]湖南省儿童医院眼科,长沙410007

出　　处：《中华实验眼科杂志》2021年第10期852-856,共5页Chinese Journal Of Experimental Ophthalmology

基　　金：湖南省科技厅计划项目(2018SK50104);爱尔眼科医院集团科研基金项目(AF1901D8)。

摘　　要：目的研究急性高眼压诱导小鼠视网膜缺血-再灌注损伤模型中血-视网膜外屏障的破坏情况。方法选取SPF级雄性C57BL/6J小鼠57只,采用随机数表法将小鼠随机分为对照组和高眼压组,其中对照组25只,高眼压组32只,剔除建模失败或建模不佳的小鼠后,每组最终纳入20只,均选取左眼为实验眼。高眼压组采用质量分数0.9%氯化钠溶液前房灌注法建立小鼠急性高眼压诱导的视网膜缺血-再灌注损伤模型,对照组仅作前房穿刺。采用动物光相干断层扫描法检测视网膜厚度变化;采用免疫荧光染色法检测视网膜组织中紧密连接蛋白1(ZO-1)分布变化;采用胰蛋白酶消化法检测视网膜毛细血管退化程度;采用苏木精-伊红染色法观察视网膜炎性细胞浸润情况。结果与对照组比较,高眼压组视网膜色素上皮(RPE)层结构紊乱,伴有明显渗出,局部神经上皮层脱离、隆起。高眼压组小鼠视网膜全层厚度为(235.8±5.3)μm,较对照组的(213.3±3.9)μm明显增厚,差异有统计学意义(t=3.427,P=0.009)。小鼠RPE层中ZO-1主要定位在细胞膜上,少部分在细胞质中,建模后2 d,高眼压组ZO-1出现明显内化,细胞膜上ZO-1减少,细胞质中ZO-1增多,整体分布紊乱。建模后7 d,高眼压组视网膜毛细血管出现退行性变,退化的毛细血管数量明显增加,高眼压组退化的毛细血管数量为(201.0±13.2)根,明显多于对照组的(11.2±1.7)根,差异有统计意义(t=14.280,P<0.01)。苏木精-伊红染色结果显示,高眼压组小鼠视网膜中可见炎性细胞浸润,主要是嗜中性粒细胞,浸润的炎性细胞主要位于内界膜下。结论急性高眼压诱导小鼠视网膜缺血-再灌注损伤使视网膜外屏障受到破坏,引起外周循环中粒细胞的浸润及视网膜水肿,视网膜毛细血管退化。Objective To investigate the breakdown of blood-retinal outer barrier in the ischemia-reperfusion injury mice following acute intraocular hypertension.Methods Fifty-seven SPF male C57BL/6J mice were selected and divided into the control group and high-intraocular pressure(IOP)group by using the random number table method.There were 25 mice in the control group and 32 mice in the high-IOP group.After the failure and poor modeling excluded,20 mice were included in each group,and the left eyes were selected as the experimental eyes.The ischemia-reperfusion injury model of the high-IOP group was established following acute intraocular hypertension by anterior chamber perfusion of 0.9%sodium chloride solution,and the control group only received anterior chamber puncture.Optical coherence tomography was used to detect retinal thickness.Immunofluorescence staining was utilized to identify zonula occludens-1(ZO-1)protein distribution in retina.Retinal capillary degeneration was identified by trypsin digestion.Inflammatory cell infiltration in retinal sections was observed by hematoxylin and eosin staining.The use and care of the animals complied with the Statement of the Association for Research in Vision and Ophthalmology,and the study protocol was approved by an Ethics Committee of Changsha Aier Eye Hospital(No.2018-KYPJ005).Results Compared with the control group,the structure of the retinal pigment epithelium(RPE)layer was irregular with obvious exudation and local neuroepithelial detachment and elevation.The thickness of the full retinal layer of mice in the high-IOP group was(235.8±5.3)|Jim,which was significantly thicker than(213.3±3.9)Rm in the control group(t=3.427,P=0.009).ZO-1 staining results showed that ZO-1 was mainly located in cell membrane and a small part in cytoplasm in the RPE layer of mice.Two days after modeling,ZO-1 in the high-IOP group was significantly internalized with decrease in cell membrane and increase in cytoplasm,and its distribution was irregular.Seven days after modeling,retinal capi

关 键 词：青光眼 高眼压 急性 血-视网膜屏障 缺血-再灌注 视网膜 病理机制 紧密连接蛋白1 

分 类 号：R775[医药卫生—眼科]