内脏脂肪素和血管内皮生长因子在早期糖尿病大鼠视网膜中的表达及意义  

Concomitant expression and combined localization of visfatin and vascular endothelial growth factor in retinas of diabetic rats

作　　者：路强 杨晓静 崔巍 高伟 

机构地区：[1]内蒙古自治区人民医院眼科,呼和浩特010017 [2]内蒙古中蒙医院眼科,呼和浩特010021

出　　处：《中华实验眼科杂志》2013年第1期45-48,共4页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目（81260152）;内蒙古自治区自然科学基金项目（2009MS1111）;内蒙古自治区人民医院科研基金项目（20110929、20110907）

摘　　要：背景糖尿病视网膜病变（DR）是糖尿病主要的微血管并发症之一，已成为糖尿病患者致盲的首要原因。新生血管形成是DR的主要病理表现，但其作用机制不明，对其发病机制的探讨有助于为DR的早期治疗提供潜在靶点。目的观察内脏脂肪素（visfatin）和血管内皮生长因子（VEGF）在糖尿病大鼠视网膜中的表达和分布特点，探讨visfatin和VEGF在DR发生发展过程中的作用机制。方法将60只雄性SPF级SD大鼠按随机数字表法分为糖尿病组和对照组，糖尿病组大鼠腹腔内一次性注射链脲佐菌素（STZ）60nlg／kg（0．60ml／100g）诱导糖尿病大鼠模型，血糖≥16．7mmol／L为造模成功，对照组以同样的方法注射等体积柠檬酸钠缓冲液。于模型建立后12周处死大鼠并提取视网膜组织，采用免疫荧光法和Westernblot法分别从形态学和分子生物学水平观察和检测visfatin和VEGF在视网膜中的表达。结果造模后12周糖尿病组大鼠平均体质量为（189．02±11．34）g，明显低于对照组的（489．57±14．48）g，差异有统计学意义（t=5．236，P=0．003）；造模后12周糖尿病大鼠血糖水平为（29．25±3．86）mmol／L，明显高于对照组的（5．32±1．01）mmol／L，差异有统计学意义（t=11．778，P=0．000）。免疫荧光双标法检测显示，对照组大鼠视网膜中visfatin和VEGF主要在视网膜神经纤维层和星形胶质细胞中呈少量表达，但在早期糖尿病大鼠视网膜全层中可见visfatin及VEGF强阳性染色，且二者呈共表达。Westernblot检测显示，造模后12周糖尿病大鼠视网膜中visfatin表达水平（A值）为346．26±41．23，对照组为102．07±65．01，差异有统计学意义（t=8．291，P=0．000）；造模后12周糖尿病组大鼠视网膜中VEGF蛋白表达量（A值）为415．88±92．15，明显高于对照组大鼠的113．06±32．06，差异有统计学意义（t=10．067，P=0．000）。Background Diabetic retinopathy （DR） is one of the most important microvascular complications of diabetes, which has become one of the leading causes of blindness. Neovascularization is the main pathological manifestations of DR,but its mechanism is unknown. There is a clear need to investigate its pathogenesis which can offer potential therapeutic targets. Objective The aim of this study was to investigate the expression and distribution of visfatin and vascular endothelial growth factor （VEGF） in diabetic model rats. Methods This study was approved by Animal Ethic Committee of Inner Mongolia Medical University. Sixty SPF 8-week-old male Sprague- Dawley rats were randomized into the diabetic group and control group. The rats were housed under a condition that alternated between 12 hours of light and darkness, with free access to rat food and water. Diabetes was induced by intraperitoneal injection of 60 mg/kg （0.60 ml/100 g） of streptozotocin （STZ） and control rats received equivalent volume of buffer. The models were regarded as successful when blood glucose was ≥ 16.7 mmol/L. Rats were sacrificed 12 weeks after the injection of STZ and retinal specimens were prepared to detect the expression of visfatin and VEGF. Total retinal protein was isolated from the retinas of experimental and control eyes,and the expression of visfatin and VEGF was assessed by Western blot. Frozen cross sections of retinas of 5 μm thickness were used to perform double immunofluorescence staining with anti-visfatin and anti-VEGF antibodies. Results Mean body weight of the diabetic rats was （189.02±11.34） g and that of the control rats was （489.57± 14.48 ） g at 12 weeks post-injection,showing a significant difference between them （ t = 5. 236, P = 0. 003 ）. Mean blood glucose level was （29.25±3.86） mmol/L in the diabetic group and （ 5.32± 1.01 ） mmol/L in the control group, demonstrating a significant difference （ t = 11. 778, P = 0. 000 ）. Double immunofluorescence staining showed reduced e

关 键 词：内脏脂肪素 血管内皮生长因子 糖尿病视网膜病变 免疫荧光双标法 免疫印迹法 

分 类 号：R77[医药卫生—眼科] R587[医药卫生—临床医学]