轴突导向因子netrin-1对早期糖尿病大鼠血视网膜屏障的影响  

The effect of netrin-1 on the retinal vascular permeability in early diabetic retinopathy rats

作　　者：周贤慧[1] 孟旭霞[1] 孙运波[1] 胡迭 郭庆敏[1] 

机构地区：[1]青岛大学附属医院眼科,266500

出　　处：《中华眼底病杂志》2015年第5期472-476,共5页Chinese Journal of Ocular Fundus Diseases

基　　金：基金项目：山东省自然科学基金（ZR2010HM062）

摘　　要：目的观察探讨外源性轴突导向因子netrin-1对早期糖尿病（DM）大鼠血视网膜屏障（BRB）的影响及可能的作用机制。方法健康雄性Sprague-Dawley大鼠80只，采用随机数字表法将其分为正常对照组、DM＋平衡盐溶液（BSS）组、DM＋netrin-1低剂量组、DM＋netrin-1高剂量组，每组均为20只大鼠。链脲佐菌素腹腔注射诱导DM模型。DM3个月时，DM＋netrin-1低剂量组、DM十netrin-1高剂量组大鼠玻璃体腔分别注射10、100mg／ml的netrin-15μl；DM＋BSS组大鼠玻璃体腔注射等体积BSS。高糖高脂饲养1个月后处死所有大鼠，摘除眼球。每组随机选取5只大鼠行苏木精-伊红染色观察各组大鼠视网膜微血管改变；免疫组织化学染色检测视网膜闭合蛋白（occludin）的表达以及荧光定量聚合酶链反应（RT—PCR）观察occludinmRNA的表达；伊凡思蓝（EB）渗漏试验观察大鼠视网膜微血管结构改变和渗漏。独立样本t检验对两组间指标进行统计学分析。多组间比较采用单因素方差分析。结果光学显微镜观察发现，DM＋BSS组大鼠视网膜神经节细胞层水肿，排列紊乱，视网膜微血管扩张，结构异常；DM＋netrin-1高剂量组大鼠视网膜神经节细胞水肿减轻，微血管扩张程度减轻。免疫组织化学染色结果显示，DM＋netrin-1低剂量组、DM＋netrin-1高剂量组大鼠视网膜棕黄色颗粒染色范围扩大，颜色加深。RT—PCR检测结果显示，与正常对照组大鼠视网膜中occludinmRNA表达比较，其余各组大鼠视网膜中occludinmRNA表达减少，差异有统计学意义（P〈0．05）。DM＋BSS组、DM＋netrin-1低剂量组、DM＋netrin-1高剂量组间大鼠视网膜平均积分吸光度[A，旧称光密度（oD）]值（IA值）比较，差异有统计学意义（F=177．13，P=0．00），且DM＋netrin-1高剂量组大鼠视网膜occludin表达较DM＋netrin-1低剂量组高。DM＋BSS组大鼠视网膜JA值与DM＋netrin-1低�Objective To explore the effect and mechanism of netrin-1 on blood-retinal barrier permeability in diabetes mellitus （DM） rats. Methods Eighty Sprague-Dawley rats were randomly divided into the normal control group, DM＋balanced salt solution （BSS） group, DM＋netrin-1 low dose group and DMq-netrin-1 high dose group, with 20 rats in each group. DM rats were induced by intraperitoneal injection of streptozocin （STZ）. These rats were feed with high sugar and fat for 3 months after STZ injection. All rats were sacrificed at i month after intravitreal injection. Retinal vascular permeability was measured by Evans blue. The expression level of oecludin was determined by immunohistochemistry. Hematoxylin-eosin （HE） staining of retina was used to observe the pathological change of DM and the levelof occludin mRNA was analyzed by real-time fluorescence quantitative reverse transcription polymerase chain reaction （RT-PCR）. Five rats of each group. Results HE staining of retina showed that the degree of edema and vascularization in DM＋ netrin-1 high dose group was better than DM-kBSS group. Staining of occludin in retina was limited to nerve fiber layer, ganglion cells, inner plexiform layer and inner nuclear layer in normal rats, but in DMq-BSS group, the color of staining positive of occludin was lighter and more reduced. However, DM＋ netrin-1 group occludin staining was deepen and enlarged. The result of RT-PCR showed that the expression of occludin mRNA in other three groups was less than normal control group （P〈0.05）. The significant difference during DM＋BSS group, low dose group and DM＋netrin i high dose group （F= 177. 13, P = 0.00）, and the more concentrate of netrin 1 the higher expression of occluding. Compared the DM＋ netrin-1 low dose group with DM＋ BSS group, there was significant difference expression of occludin （t=- 13.98, P=0.00）. There was significant difference between the DM＋netrin 1 high dose group and normal control group （t = 12.87, P = 0.00）. There

关 键 词：糖尿病视网膜病变/病理生理学 血视网膜屏障/病理生理学 神经生长因子类 动物实验 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]