糖尿病大鼠视网膜基因表达谱差异的初步分析

Preliminary analysis of retinal gene expression profile of diabetic rat

目的建立大鼠正常视网膜和糖尿病8周视网膜基因表达谱,比较两者差异,初步分析糖尿病视网膜病变的相关基因。方法通过限制片段差异显示PCR(restriction fragments differential display-PCR,RFDD-PCR)获得正常大鼠视网膜及8周糖尿病大鼠视网膜组织转录组片段。应用Fragment Analysis等软件,对差异片段进行生物信息学分析,初步确定糖尿病视网膜病变相关基因/表达序列标签(expression se-quence tag,EST)。结果获得有意义的片段共3639个,有差异的片段840个,占表达数的23.08%。其中包括5个视觉传导相关基因,13个兴奋性神经递质受体基因和3个抑制性神经递质受体基因。糖尿病8周大鼠视网膜Rhodopsin kinase、β-arrestin、Phosducin、rod photoreceptor cGMP-gated channel和Rpe65的表达下调,离子型谷氨酸受体iGluR1-4下调,代谢性谷氨酸受体及γ-氨基丁酸受体各亚型则普遍上调,而甘氨酸受体表达无变化。结论糖尿病8周大鼠神经视网膜已受到累及,其基因表达模式的改变,可能与糖尿病早期视功能损害有关。

Objective Establishing the retinal gene expression profiles of non-diabetic rat and diabetic rat and comparing the profiles in order to analyze the possible genes related with diabetic retinopathy, nelhods The whole retinal transcriptional fragments of non-diabetic rat and 8-week diabetic rat were obtained by restriction fragments differential diaplay-PCR （RFDD-PCR）. Bioinformatic analysis of retinal gene expression was performed using soft wares, including Fragment Analysis. After comparison of the expression profiles, the related gene fragments of diabetic retinopathy were initially selected as the target gene of further approach. Results A total of 3639 significant fragments were obtained. By means of more than 3-fold contrast of fluorescent intensity as the differential expression standard, the authors got 840 differential fragments, accounting for 23.08% of the expressed numbers and including 5 visual related genes, 13 excitatory neruotransmitter genes and 3 inhibitory neurotransmitter genes. At the 8th week, the expression of Rhodopsin kinase, β-arresfin, Phosducin, rod photoreceptor cGMP-gated channel and Rpe65 as well as iGlu R1-4 were down-regulated. mCduRs and GABA-Rs were all up-regulated, whereas the expression of GlyR was unchanged. Conduslon These resuits prompt again that the changes in retinal nervous layer of rat have occurred at an early stage of diabetes. The genes expression pattern of visual related genes and excitatory and inhibitory neurotransmitters in rat diabetic retina have been involved in neuro-dysfunctions of diabetic retina.