多药耐药蛋白1和多药耐药相关蛋白5、7在糖尿病小鼠视网膜中表达的变化  

Differential expression of MDR1,MRP5 and MRP7 in diabetic mice retina

作　　者：李梦双[1,2] 郭传龙[2] 李君[2] 吴祥根[1,2] 

机构地区：[1]济南大学山东省医学科学院医学与生命科学学院,山东省济南市250022 [2]山东省眼科研究所,山东省青岛市266071

出　　处：《眼科新进展》2017年第1期10-14,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:81271036;81500751);2015年度山东省重点研发计划资助(编号:2015GSF11-8121);山东省医学科学院医药卫生科技创新工程~~

摘　　要：目的观察多药耐药蛋白1(multidrug resistance 1,MDR1)、多药耐药相关蛋白(multidrug resistance-associated protein,MRP)5和MRP7在不同时期糖尿病小鼠视网膜中的表达变化。方法通过腹腔注射链脲佐菌素诱导C57BL/6小鼠建立糖尿病模型,伊文思蓝灌注检测血-视网膜内屏障变化。分别于糖尿病造模成功后的4周、12周和24周取小鼠视网膜,进行实时定量-PCR、免疫荧光、Western blotting等实验,检测各组小鼠视网膜中MDR1、MRP5和MRP7的表达变化。结果伊文思蓝灌注结果显示在糖尿病12周就出现视网膜渗漏;实时定量-PCR结果发现与对照组相比,随糖尿病进展,MDR1、MRP5和MRP7在4周、12周和24周的表达均呈下降趋势(MDR1:P_(4周)=0.028,P_(12周)=0.003,P_(24周)<0.001;MRP5:P_(4周)=0.045,P_(12周)=0.009,P_(24周)<0.001;MRP7:P_(4周)=0.019,P12周<0.001,P24周=0.001),在糖尿病不同时期其变化也呈下降趋势(均为P<0.05);免疫荧光观察到MDR1、MRP5和MRP7的荧光强度随糖尿病发展逐渐减弱;Western blotting检测可见在4周、12周、24周糖尿病小鼠视网膜中MDR1、MRP5和MRP7的表达均呈下降趋势。结论在DR早期血-视网膜内屏障会受到破坏,视网膜中外排转运蛋白的表达也会受糖尿病的影响导致不同程度的下降。Objective To investigate the expression of multidrug resistance 1 （MDR1）, multidrug resistance-associated protein 5 （ MRP5 ）, and multidrug resistanceassociated protein 7 （ MRP7 ） in the diabetic mice retina during different periods. Methods The C57BL/6 mice underwent induction of diabetes mellitus with an intraperitoneal injection of streptozotocin. The changes of blood-retinal barrier were detected by perfusion with Evans blue. RT-PCR, immunofluorescence, and Western blotting were used to obtain the changes of MDR1, MRPS, and MRP7 in the diabetic mice retina at 4 weeks,12 weeks,and 24 weeks. Results The results of Evans blue perfusion showed that the retinal vascular leakage was occurred at 12 weeks. RT-PCR showed that the expression of MDR1 ,MRP5 ,and MRP7 were all declined with the development of diabetic when compared with control group （MDR1 ：P4 weeks = 0. 028, P,2 weeks = 0. 003, P24 weeks 〈 0. 001 ;MRP5 ：P4 weeks = 0. 045, P12 weeks = 0. 009, P24 weeks 〈 0. 001 ; MRP7 ： P4 weeks = 0. 019, P12 weeks 〈 0. 001 ,P24 weeks = 0. 001 ）, and the trend of changes were decreased among different period of diabetes meUitus（ all P 〈 0.05 ）. The immunofluorescence observed that the fluorescence intensity of MDR1,MRPS, and MRP7 were decreased. As for the West- ern blotting, the expression of MDR1, MRPS, and MRP7 were down-regulated in diabetic mice retina. Conclusion The blood-retinal barrier is disrupted in early diabetic retinopathy, and the expression of efflux transporters in retina are decreased due to diabetes mellitus.

关 键 词：糖尿病视网膜病变 血-视网膜内屏障 多药耐药蛋白1 多药耐药相关蛋白5 多药耐药相关蛋白7 

分 类 号：R774.1[医药卫生—眼科]