新生小鼠视网膜病变的基因表达谱变化

Change of gene expression profile and its significance in mouse with oxygen-induced retinopathy

目的应用基因芯片技术研究早产儿视网膜病(retinopathy of prematurity,ROP)基因表达谱的变化规律及意义。方法7日龄C57BL/6J新生小鼠暴露于75%的高氧5d后回到空气中,制成ROP动物模型,应用Trizol法抽提17日龄小鼠ROP模型及同时期对照组小鼠的视网膜总RNA,逆转录并用Cy3和Cy5标记,与小鼠Oligo表达谱芯片杂交,扫描后获取基因表达信息,进行差异分析、聚类分析和基因分类等生物信息学分析。结果与正常小鼠比较,视网膜病变小鼠有1347个基因呈差异表达。SOM聚类显示12类不同的表达差异变化模式,基因本体学分析显示变化的基因并不局限于血管生成相关因子,发育相关基因及G蛋白信号通路相关基因改变亦很明显(P<0.05)。结论ROP小鼠视网膜基因表达谱有较大改变,为今后从分子生物学角度探索该病发病机制和进行基因治疗奠定基础。

Objective To study the change of gene expression profile and its significance in mouse with oxygen-induced retinopathy by gene chips. Methods ROP model was developed in C57BL/6J mice by 5-day exposure to 75% oxygen from 7 days after born （P7） to P12 followed by 5- day and back to room air. Pooled total RNA was prepared with a Trizol method from retinae of the following two groups： hyperoxic ROP group and control group with matched age. Purified mRNA was reverse transcripted to cDNA, and labelled with Cy3 and CyS. Labelled cDNA was hybridized with an expression profile microarray of oligonucleotides of mouse genes. Signals were extracted from the scanned images of chips, followed by background extraction and normalization, and subjected for further analysis, such as ratio, cluster and gene ontology（GO） analysis. Results There were 1347 differentially expressed genes in mice with retinopathy of prematurity, in which 646 genes were upregulated and 378 genes were downregulated by more than 2 times. These genes were clustered to 12 groups with different expression patterns according to self-organizing map（SOM） method. GO analysis indicated that differentially expressed genes were not limited to vascular growth associated factors, but genes in correlation to organogenesis and G-protein signaling pathway had significant changes（P〈0. 05）. Conclusions The change of global gene expression profile in retinae of mouse with retinopathy of prematurity was obvious.