cDNA基因芯片技术检测妊娠合并糖尿病诱发胚胎先天性神经管缺陷表达差异基因(英文)

Identification of differentially expressed genes involved in diabetes-induced embryopathy by cDNA microarray

目的探讨妊娠合并糖尿病诱发胚胎先天性神经管缺陷的差异表达基因,揭示胚胎先天性神经管缺陷发生、发展的分子机制。方法选用2组的70～90天的SD大鼠:第1组(n=3)为接受常规饲养的正常对照组;第2组(n=3)为实验组:尾静脉注射65mg/kg链脲菌素(STZ),构建妊娠合并糖尿病诱发胚胎先天性神经管缺陷组大鼠动物模型。于妊娠第12天取出各组胚胎,解剖显微镜下进行神经管缺陷形态学判定。提取卵黄囊细胞的mRNA,以cDNA基因芯片技术对差异表达基因进行检测。结果对实验组及正常对照组卵黄囊细胞的1200个基因进行比较,共筛选出差异表达基因79条,其中42条表达上调基因(包括凋亡相关基因BAX、bcl 2、HSP70、glucose transporter3等),37条表达下调基因(包括phospholipaseA2、insulin likegrowthfactorIIreceptor等)。结论揭示了妊娠合并糖尿病诱发胚胎先天性神经管缺陷的差异表达基因,对先天性神经管缺陷有效的早期诊断和治疗提供了实验依据。

Objective Our purpose in this study is to investigate genes involved in the development of diabetes-induced embryonic malformations. Methods Two groups of 70-90 day old Sprague-Dawley rats were employed in our study: group 1 was normal control rats receiving a normal diet (n=3); group 2 consisted of experimentally-induced diabetic rats by intravenous injection of 65mg/kg of streptozotocin(STZ) on pregnancy day 6 with an attempt to reproduce malformations in embryos (n=3). Embryos were examined on day 12 under light microscopy to look for morphological defect of the neural tube (NTD). Yolk sac cells were harvested from each group and RNA was isolated. Genes expression profiles in yolk sac cells were analyzed using a DNA microarray technique. Results Gene expression patterns were compared in a total of 1200 genes between experimentally-induced diabetic rats and normal control rats, and 79 of genes were found to express differently between the two groups. Forty-two of genes were up-regulated in yolk sac cells of diabetic rats, such as apoptosis related genes BAX, bcl-2, heat shock 70kD protein and glucose-transporter 3; 37 of genes were down-regulated, such as phospholipase A 2, insulin-like growth factor II receptor. Conclusion Understanding of differently expressed genes should help us disclose the potential molecular mechanisms underlying the developmental process during diabetes-associated embryonic morphogenesis, and it also might provide a useful tool in rapid diagnosis and prevention of malformation in early gestation stage of diabetic subjects.