胰岛素样生长因子结合蛋白-6在增生性玻璃体视网膜病变大鼠玻璃体和血清中的表达  

Expression of insulin-like growth factor binding protein-6 in the vitreous and serum in a proliferative vitreoretinopathy rat model

作　　者：于靖[1] 崔尘[2] 赵红梅[3] 王克生[4] 

机构地区：[1]同济大学附属第十人民医院眼科,上海200072 [2]南通大学附属医院眼科 [3]复旦大学附属眼耳鼻喉医院眼科 [4]复旦大学中心实验室

出　　处：《中华实验眼科杂志》2013年第1期65-69,共5页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金青年基金项目（30901643）;上海市科委生物医药处医学引导项目（10411964900）;上海市卫生系统新优青计划项目（XYQ2011067）;上海市卫生局基金项目（2008179）

摘　　要：背景增生性玻璃体视网膜病变（PVR）是导致视网膜脱离复位手术失败的主要原因之一。PVR玻璃体蛋白质组学研究发现，胰岛素样生长因子结合蛋白-6（IGFBP-6）是PVR患者玻璃体和血清中的特异蛋白质，且其在严重PVR患者中的含量明显高于轻度PVR。目的探讨IGFBP-6在PVR大鼠模型中的表达变化。方法选取7周龄SPF级雄性Wistar大鼠70只，采用抛硬币法随机将动物分为PVR模型组34只和对照组36只。PVR模型组大鼠左眼玻璃体腔内注入透明质酸酶1U（商品单位），注射后10min待玻璃体液化后，玻璃体腔内再注入1×106视网膜色素上皮（RPE）细胞悬液5μl和1×107富含血小板的血浆5μl；对照组大鼠以同样的方法注射无菌生理盐水10μl。各组大鼠玻璃体腔注射后1、2、3、4周行眼底检查并按Francine标准对PVR进行分级。分别于造模后1、2、4、8周处死各组大鼠，抽取动物心脏血，切取肝脏和视网膜，并制备切片，采用实时荧光定量PCR法检测大鼠肝脏组织和视网膜组织中IGFBP-6mRNA的表达，采用ELISA法检测大鼠玻璃体和血清中IGFBP-6的质量浓度，对各组大鼠的测量指标进行比较。结果利用实时荧光定量PCR法分别从肝脏和视网膜中提取高纯度的IGFBP-6mRNA。PVR模型组大鼠造模后第4周视网膜中IGFBP-6mRNA含量为（3．79±1．33）×10-4，明显低于对照组的（8．32±2．96）×10-4，差异有统计学意义（t=3．420，P〈0．01）；PVR模型组大鼠造模后第4周视网膜中IGFBP-6mRNA含量明显低于造模后第1、2、8周，差异均有统计学意义（P〈0．05），而PVR模型组大鼠造模后各时间点肝脏中IGFBP-6mRNA含量与对照组比较以及PVR模型组大鼠组内各时间点间的比较差异均无统计学意义（P〉0．05）。PVR1、2、3级组大鼠视网膜中IGFBP-6mRNA含量明显低于对照组，差异有统计学意义（P〉0．05），但不同等级的PVR大鼠间差异无�＇Abstract] Background Proliferative vitreoretinopathy （PVR） is one of the major causes of retinal detachment surgery failure. Based on proteomic studies of PVR vitreous,the insulin-like growth factor binding protein-6 （IGFBP-6） protein was specifically expressed in the vitreous and serum of PVR patients. Furthermore,its expression level is higher in the vitreous and serum in severe PVR patients than that in mild PVR patients. Objective This experiment was to detect the expression of IGFBP-6 in a PVR rat model. Methods Seventy 7-week old male SPF Wistar rats were included and were randomized into the PVR model group and control group. A mixture of RPE-J cell suspension（5 μl） and platelet-rich plasma （5 μl） was intravitreally injected in the left eyes of adult Wistar rats to establish the PVR model,and normal saline solution was administered in the same way in the control group. The rat eyes were clinically examined 1 week,2,3 and 4 weeks after injection, and PVR was graded based on the criteria of Franeine. The animals were sacrificed after 1 week,2,4 or 8 weeks for the preparation of retinal sections and liver extraction. Expression levels of IGFBP-6 mRNA in the rat retina and liver were assayed by real-time Q-PCR. The expression of IGFBP-6 protein in the rat serum and vitreous was detected by ELISA. The use of animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission. Results Purified IGFBP-6 RNA was extracted from the liver and retina of Wistar rat and quantified by real-time Q-PCR. The expression level of IGFBP-6 mRNA in retina was（3.79±1.33 ）×10-4 in the PVR model rats, showing a significant decline in comparison with the control rats with a level of（8.32±2.96 ）×10-4,4 weeks after injection （t = 3.42, P〈0. 01 ）. The expression of IGFBP-6 mRNA in the 4th week was significantly lower than that of 1 week,2 or 8 weeks after the establishment of the PVR model（P〈0.05）. No significant

关 键 词：胰岛素样生长因子结合蛋白 增生性玻璃体视网膜病变 大鼠 模型 视网膜 

分 类 号：R77[医药卫生—眼科]