胚胎先天性神经管缺陷的表达差异基因及信号传导  

Identification of differentially expressed genes and signal pathways involved in diabetes-induced congenital neural tube defects

作　　者：马向东[1] 吴小明[2] 马兴[3] 赵东涛[2] 陈必良[1] 辛晓燕[1] 王德堂[1] 

机构地区：[1]第四军医大学西京医院妇产科,陕西西安710033 [2]第四军医大学生物医学工程系 [3]第四军医大学西京医院骨科,陕西西安710033

出　　处：《第四军医大学学报》2008年第19期1737-1740,共4页Journal of the Fourth Military Medical University

基　　金：国家自然科学青年基金(30200297)

摘　　要：目的:研究妊娠合并糖尿病诱发胚胎先天性神经管缺陷的分子机制.方法:将29只SD雌性大鼠分为3组:正常对照组,8只;②实验模型组,链脲佐菌素(STZ)构建的妊娠合并糖尿病诱发胚胎先天性神经管缺陷,12只;③实验对照组,STZ构建的糖尿病大鼠模型但胚胎不伴有先天性神经管缺陷发生,9只.于妊娠第12日取出各组胚胎,解剖显微镜下进行形态学分析.提取卵黄囊细胞mRNA,用cDNA基因芯片技术对表达差异基因进行检测.提取卵黄囊细胞蛋白质,应用特异性抗磷酸化抗体进行免疫共沉淀及WesternBlot,对MAP Kinase信号途径上各蛋白激酶活性进行分析.结果:提取实验模型组大鼠胚胎卵黄囊细胞mRNA,与正常对照组大鼠胚胎卵黄囊细胞mRNA共1200个基因同时进行比较,共筛选出表达差异基因79条,其中42条表达上调基因,包括凋亡相关基因BAX,bcl-2,heat shock70,glucose-transporter3等;37条表达下调基因包括phospholipase A2,insulin-likegrowth factorⅡreceptor等.与正常对照组相比较,ERK1/2蛋白激酶活性显著下降;JNK1/2活性明显升高,凋亡相关蛋白Caspase-3,Bax高表达,凋亡抑制蛋白AKT活性明显受抑.结论:妊娠合并糖尿病诱发的胚胎先天性神经管缺陷的发生,与MAP Kinase及细胞凋亡信号传导机制参与密切相关.AIM： To determine the molecular mechanism in hyperglycemia-induced congenital neural tube defects. METHODS： Three study groups of Sprague-Dawley rats were employed： Group 1 were 8 normal control rats with normal diet; group 2 represented 12 streptozotocin（STZ）-induced diabetic rats with congenital neural tube defects in offspring; group 3 included 9 STZ-induced diabetic rats with normal offspring. Yolk sac cells were harvested at day 12 of gestation from each rat group, and mRNA was isolated. We analyzed gene expression profiles in yolk sac cells using a cDNA microarray technique. Changes in MAPK and apoptosis signaling pathways were detected by Western Blot analyses using special antibodies. RESULTS： In comparison of gene expression patterns between experimentally-induced diabetic rats and normal control rats in total 1200 genes, 79 genes were differentially expressed between two groups. Fourty-two genes were up-regulated in yolk sac cells of diabetic rats with congenital neural tube defects, such as apoptosis related genes BAX, bcl-2, heat shock 70-kDa and glucose-transporter 3; and 37 genes were down-regulated, such as phospholipase A2, insulin-like growth factor Ⅱ receptor. Activities of ERK1/2 were dramatically decreased （group 2）. In contrast, activities of JNK1/2 were significantly increased in yolk sac cells of group 2. Similarly, increased expressions of pro-apoptotic caspase-3 and Bax genes were observed in yolk sac cells in the same group; whereas phosphorylated AKT was down-regulated. CONCLUSION： Understanding differentially expressed genes would help us to disclose the potential molecular mechanisms in the developmental process of diabetes-associated embryonic morphogenesis. MAP kinase and apoptotic signal pathways play very important roles in the development of hyperglycemia induced neural tube defects.

关 键 词：CDNA 基因芯片 神经管缺陷 糖尿病 MAP KINASE 细胞凋亡 

分 类 号：R726.2[医药卫生—儿科]