糖尿病大鼠视网膜组织突触核蛋白家族的表达  

Expression of synuclein proteins family in retinal tissue of diabetic rats

作　　者：袁爱花[1] 梅妍[2] 周鸿鹰[2] 羊惠君[2] 

机构地区：[1]上海交通大学医学院附属精神卫生中心遗传研究室,上海市200030 [2]四川大学华西医学中心基础医学院人体解剖与组织胚胎学教研室,四川省成都市610041

出　　处：《眼科新进展》2012年第3期215-218,共4页Recent Advances in Ophthalmology

基　　金：2010年上海市卫生局青年基金资助(编号:2010Y030)~~

摘　　要：目的观察突触核蛋白(synuclein)家族三种同源蛋白基因α-synuclein、β-synucle-in和γ-synuclein在8周糖尿病大鼠视网膜组织中的表达情况,初步探讨其在糖尿病视网膜病变(diabetic retinopathy,DR)中的作用。方法采用限制性片段差异显示聚合酶链反应技术,建立正常和8周糖尿病大鼠视网膜基因表达谱,经生物信息学分析两者差异,初步确定α-synuclein、β-synuclein和γ-synuclein为DR相关基因,并以实时定量PCR和Western blot-ting方法分别从mRNA和蛋白水平上对三个基因的表达进行验证。结果限制性片段差异显示聚合酶链反应结果显示,糖尿病大鼠α-synuclein、β-synuclein和γ-synuclein表达明显强于正常大鼠;实时定量PCR结果显示,α-synuclein、β-synuclein和γ-synuclein在糖尿病大鼠(1.41±0.14、1.30±0.18、1.47±0.15)的表达(相对Ct比值)高于正常大鼠(1.67±0.12、1.62±0.20、1.86±0.26),差异均有显著统计学意义(均为P<0.01);Western blotting显示,糖尿病大鼠三种蛋白的表达(相对A值比值)分别为0.31±0.06、0.60±0.11、0.39±0.09,显著高于正常大鼠(0.19±0.07、0.44±0.09、0.29±0.08),差异均有显著统计学意义(均为P<0.01)。结论α-synuclein、β-synuclein和γ-sy-nuclein在糖尿病大鼠视网膜组织中表达显著升高,α-synuclein的表达升高可能在DR中起到神经毒性作用,而β-synuclein和γ-synuclein的高表达则可能对神经细胞起到保护作用。Objective To observe the expression of three kinds of homologous proteins of synuclein protein family including α-,β-and γ-synuclein in the retina of 8-week diabetic rat,and explore the potential roles of α-,β-and γ-synuclein in diabetic retinopathy.Methods Retinal gene expression profiles of healthy and 8-week-old diabetic rats were established with restriction fragment differential display polymerase chain reaction（RFDD-PCR）,and the differences of the two profiles were analyzed with bioinformatics,α-,β-and γ-synuclein were identified as the related gene of DR,and the levels of mRNA and proteins of α-,β-and γ-synuclein were verified with fluorescence quantitative PCR and Western blotting.Results The results of RFDD-PCR showed the expression of α-,β-and γ-synuclein in diabetic retina of rats was significantly stronger than those in the healthy retina of rats.The results of fluorescence quantitative PCR showed that the mRNA levels（relative ratio of Ct） of α-,β-and γ-synuclein in diabetic retina（1.41±0.14,1.30±0.18,1.47±0.15） were much higher than those in healthy retina of rats（1.67±0.12,1.62±0.20,1.86±0.26）,the expression difference had statistical significances（all P0.01）;The results of Western bloting showed that the protein levels（relative ratio of A） of α-,β-and γ-synuclein in diabetic retina（0.31±0.06,0.60±0.11,0.39±0.09） were much higher than those in healthy retina of rats（0.19±0.07,0.44±0.09,0.29±0.08）,the expression difference had statistical significances（all P0.01）.Conclusion The expression level of α-,β-and γ-synuclein in the retina of diabetic rat is significant high.The enhanced expression of α-synuclein perhaps has neurotoxic effects,while the enhanced expression of both β-synuclein and γ-synuclein may have neuroprotective effects in diabetic retinopathy.

关 键 词：糖尿病视网膜病变 突触核蛋白 基因表达谱 定时定量PCR WESTERN BLOTTING 

分 类 号：R587.1[医药卫生—内分泌]