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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 To establish the retinal gene expression profiles of normal retina and diabetic rats for 8 weeks, and to compare the differences between the two. The preliminary analysis of the related genes of diabetic retinopathy. Methods Retinal tissue transcriptome of normal rat retina and 8-week diabetic rats were obtained by restriction fragment differential display-PCR (RFDD-PCR). Fragment Analysis and other softwares were used to analyze the differential fragments for bioinformatics analysis, and to initially identify the expression of ESTs / ESTs. Results A total of 3639 meaningful fragments were obtained, with 840 fragments, accounting for 23.08% of the total. These include five visual transduction related genes, 13 excitatory neurotransmitter receptor genes and three inhibitory neurotransmitter receptor genes. The expression of cGMP-gated channel and Rpe65 in retina of diabetic rats after 8 weeks of treatment was down-regulated, and iGluR1-4, a metabotropic glutamate receptor, and γ-amino Butyric acid receptor subtypes were generally up-regulated, while no changes in glycine receptor expression. Conclusion The diabetic retina has been implicated in the retina of rats for 8 weeks. The changes of its gene expression pattern may be related to the impairment of early diabetic function.