Nogo蛋白受体(NgR)基因沉默对糖尿病大鼠视网膜神经节细胞密度及突触素表达的影响  

Effects of inhibiting NgR on retinal ganglion cells density and synaptophysin expression of diabetic rats

作　　者：刘文强 王玉波 梁汇珉 李赵伟 李铮 刘学政 

机构地区：[1]锦州医科大学基础医学院解剖学教研室,辽宁省锦州市121001

出　　处：《眼科新进展》2017年第2期106-109,共4页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:81571383)~~

摘　　要：目的探讨抑制Nogo蛋白受体(nogo receptor,NgR)对糖尿病大鼠视网膜神经节细胞(retinal ganglion cell,RGC)密度及突触素表达的影响。方法取雄性SD大鼠32只,随机分成正常对照组、糖尿病组、siNgR组及scN gR组,每组8只。后3组腹腔注射链脲佐菌素(50 mg·kg^(-1)),72 h后尾静脉血糖大于16.7 mmol·L-1的大鼠定为糖尿病模型。siNgR组玻璃体给予腺病毒包装的NgR反义核苷酸序列10μL,scN gR组玻璃体给予NgR阴性核苷酸序列10μL,正常对照组及糖尿病组不进行处理。8周后,HE染色检测RGC密度,免疫荧光技术检测视网膜突触素表达及分布,Western blot检测视网膜NgR及突触素相对表达量。结果正常对照组、糖尿病组、siNgR组及scN gR组RGC密度分别为(624.33±3.51)个·mm-2、(420.00±2.65)个·mm-2、(621.67±1.53)个·mm-2、(416.67±2.52)个·mm-2。4组间相比,总体差异有统计学意义(F=5985.37,P<0.01)。与正常对照组相比,糖尿病组及scN gR组RGC密度均明显降低(均为P<0.01),而siNgR组无明显变化(P>0.05)。突触素主要表达于视网膜内网层和外网层,与正常对照组相比,糖尿病组及scN gR组阳性表达减少,荧光强度减弱。而siNgR组无明显变化。正常对照组、糖尿病组、siNgR组及scN gR组NgR蛋白相对表达量分别为:(11.26±0.02)%、(19.38±0.10)%、(11.17±0.02)%、(19.47±0.31)%。4组间比较,总体差异有统计学意义(F=2466.09,P<0.01);与正常对照组相比,糖尿病组及scN gR组蛋白相对表达量均增加(均为P<0.01),而siNgR组无明显变化(P>0.05)。突触素蛋白相对表达量分别为:(35.76±0.15)%、(25.47±0.36)%、(35.28±0.12)%、(25.03±0.75)%。4组间比较,总体差异有统计学意义(F=583.70,P<0.01);与正常对照组相比,糖尿病组及scN gR组蛋白相对表达量均降低(均为P<0.01),而siNgR组无明显变化(P>0.05)。结论抑制糖尿病大鼠视网膜NgR表达有助于恢复突触数量,保护受损的RGC。Objective To investigate the effects of inhibiting NgR on retinal ganglion cells density and synaptophysin expression of diabetic rat.Methods Thirty-two SD male rats were randomly divided into normal control group,diabetic group,siNgR group and scNgR group,8rats in each group.Normal control group was given no any treatment.Diabetes model was induced by intraperitoneal injection of 50 mg · kg^（-1） streptozotocin in diabetic group,siNgR group and scNgR group,and the blood glucose more than 16.7 mmol · L^（-1） at 72 hours was set as the successfully model.The rats of siNgR group were intravitreally administrated with anti-NgR nucleotide and the rats of scNgR group intravitreally administrated with negative nucleotide.Eight weeks later,HE staining was conducted to detect density of retinal ganglion cell（RGC）,immunofluorescence was used to evaluate the expression and distribution of synaptophysin（a marker of synaptic number）.Relative expression of NgR and synaptophysin in retina were analyzed by Western blot.Results RGC density in normal control group,diabetes group,siNgR group and scNgR group were（624.33±3.51）mm^（-2）,（420.00±2.65） mm^（-2）,（621.67±1.53） mm^（-2）,（416.67±2.52） mm^（-2） respectively.There was significant difference among four groups（F =5985.37,P〈0.01）.Compared with normal control group,RGC density in diabetes group and scNgR group were obviously decreased（all P〈0.01）,but siNgR group had no obviously change（P0.05）.The synaptophysin mainly expressed in the inner and outer network layer.Compared with normal control group,the positive expression of synaptophysin in diabetes group and scNgR group were decreased,but siNgR group had no obviously change.The relative expression of NgR in normal control group,diabetes group,siNgR group and scNgR group were（11.26±0.02）%,（19.38±0.10）%,（11.17±0.02）%,（19.47±0,31）%,respectively.There was significant difference among four groups（F = 2466.09,P〈0.01）.Compared with normal control

关 键 词：糖尿病视网膜病变 视网膜神经节细胞 Nogo蛋白受体 突触素 

分 类 号：R774[医药卫生—眼科]