不同浓度外源性轴突导向因子-1对糖尿病大鼠视网膜血管通透性的影响  

The effect of netrin-1 on the retinal vascular permeability in diabetes mellitus rats

作　　者：周贤慧 孟旭霞[1] 付浴东[1] 刘鹏辉[1] 胡迭 

机构地区：[1]青岛大学附属医院眼科,266500 [2]烟台业达医院眼科 [3]青岛眼科医院

出　　处：《中华眼底病杂志》2017年第3期286-289,共4页Chinese Journal of Ocular Fundus Diseases

摘　　要：目的 观察不同浓度外源性轴突导向因子（netrin）-1对糖尿病（DM）大鼠视网膜血管通透性的影响。 方法 健康清洁级雄性Sprague-Dawley大鼠80只，采用随机数字表法随机分为正常对照组（A组）、正常＋平衡盐溶液（BSS）组（B组）、正常＋netrin-1 500 μg/ml组（C组）、DM＋BSS组（D组）、DM＋netrin-1 10 μg/ml组（E组）、DM＋netrin-1 50 μg/ml组（F组）、DM＋netrin-1 100 μg/ml组（G组）、DM＋netrin-1 500 μg/ml组（H组），每组10只。E～H组大鼠腹腔注射链脲佐菌素诱导DM动物模型。建模3个月后，E～H组大鼠玻璃体腔分别注射10、50、100、500 μg/ml netrin-1 5 μl。C组大鼠玻璃体腔注射500 μg/ml netrin-1 5 μl。B、C组大鼠玻璃体腔注射等体积BSS。A组大鼠不做任何处理。采用免疫组织化学染色法观察大鼠视网膜闭合蛋白（occludin）的阳性表达；荧光定量聚合酶链反应检测大鼠视网膜occludin mRNA的表达；伊凡思蓝检测各组大鼠视网膜血管通透性变化。 结果 与A组比较，D组大鼠视网膜occludin阳性表达（t=27.71，P=0.00）、mRNA表达（t=8.59，P=0.00）均减少，差异有统计学意义；视网膜血管通透性增加，差异有统计学意义（t=？42.72，P=0.00）。D～H组大鼠视网膜occludin阳性表达（F=146.31，P=0.00）、mRNA表达（F=16.54，P=0.00）及视网膜血管通透性（F=67.77，P=0.00）比较，差异有统计学意义。B、C组大鼠视网膜occludin阳性表达（t=？1.13，P=0.27）、mRNA表达（t=0.93，P=0.36）及视网膜血管通透性（t=1.04，P=0.31）比较，差异无统计学意义。随着外源性netrin-1浓度升高，大鼠视网膜occludin阳性表达、mRNA表达升高，视网膜血管通透性降低。相关性分析结果显示，外源性netrin-1浓度与视网膜occludin阳性表达、mRNA表达呈正相关（r=0.73、0.81，P=0.00、0.00），与视网膜血管通透性呈负相关（r=–0.61，P=0.00）。 结论 外源性nObjective To observe the effect of different concentration netrin-1 on retinal vascular permeability in diabetes mellitus （DM） rats. Methods Eighty adult Sprague-Dawley rats were randomly divided into 8 groups, 10 rats in each group, including normal control group （group A）, normal＋balanced salt solution （BSS） group （group B）, normal＋netrin-1 （500 μg/ml） group （group C） and DM group （50 rats in 5 sub-groups）. DM rats were induced by intraperitoneal injection of streptozocin. Three months after intraperitoneal injection, 10 DM rats in the control group were injected with BSS （group D）. Forty DM rats were injected with 5 μl of different concentrate netrin-1, and were divided into DM＋netrin-1 10 μg/ml group （group E）, DM＋netrin-1 50 μg/ml group （group F）, DM＋netrin-1 100 μg/ml group （group G）, DM＋netrin-1 500 μg/ml group （group H） according to the different concentration. Non-DM rats in group C were injected with netrin-1 500 μg/ml. The expression of occludin was determined by immunohistochemistry for protein, and by real-time fluorescence quantitative reverse transcription polymerase chain reaction for mRNA level. Retinal vascular permeability was measured by Evans blue infusion. Results The expression of occludin protein and mRNA in group D were less than group A （t=27.71, 8.59;P=0.00, 0.00）. However, the retinal vascular permeability increased in group D （t=？42.72,P=0.00）. The expression of occluding protein, occludin mRNA and retinal vascular permeability showed significant differences between group D, E, F, G and H （F=146.31, 16.54, 67.77;P=0.00, 0.00, 0.00）. Compared the group B with group C, there was no significant differences between the expression of occludin protein, occludin mRNA and the retinal vascular permeability （t=？1.13, 0.93, 1.04;P=0.27, 0.36, 0.31）. The concentrate of netrin-1 showed a significant positive correlation to the expression level of occludin and occludin mRNA （r=0.73, 0.81;P=0.00, 0.00）, but ne

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]