宫内糖尿病环境致子代胰岛素通路蛋白FoxO1改变及影响印记基因表达的分子机制研究

The pathogenesis of diabetes is complicated.Study the pathogenesis of fetal-derived diabetes from gestatioanal diabetes mellitus (GDM) with intrauterine environment exposure is a major issue of this research areas. The preliminary study of our group found that: Intrauterine exposure of diabetic environment of GDM can induce the alteration of important imprinted genes Igf2/H19 expression and the imprinting control region methylation abnormalities..The project intends on detecting the expression level of insulin pathway pretein FoxO1 and C/ERBa,methyl transferase 3a(Dnmt3a) and Igf2/H19, and the DMR methylation level of Igf2/H19 in the offspirng liver cells, by using of the GDM mouse model.We intends to clarify the regulation of the imprinted genes by intrauterine diabetic environment is through the insulin pathway proteins affect methyltransferase (Dnmts);The application of transfection and SiRNA technology to correct abnormal FoxO1 expression using in vitro cell culture,is used to observe the subsequent imprinted gene expression; reveal the possible mechanism of the GDM intrauterine environment increase the susceptibility of diabets in the offspring, and provide a theoretical basis of the control of fetal-derived diabetes.

糖尿病发病机制复杂，从妊娠糖尿病（GDM）的宫内环境暴露研究胎儿源性糖尿病的发病机制是该研究领域的崭新重大课题。课题组前期研究发现：GDM宫内糖尿病环境可诱发子代糖尿病发病相关重要印记基因Igf2/H19表达和印记控制区甲基化异常。本项目拟在此基础上，利用GDM小鼠模型，检测子代肝细胞胰岛素通路蛋白FoxO1、甲基转移酶3a（Dnmt3a）和印记基因Igf2/H19表达水平以及Igf2/H19 DMR甲基化水平，阐明宫内糖尿病环境对印记基因的调控是通过胰岛素通路蛋白影响甲基转移酶所致；进行体外细胞培养，应用转染和SiRNA技术，纠正异常FoxO1表达，比较分析改善胰岛素通路调节异常对后续印记基因表达异常的消减作用；揭示GDM宫内糖尿病环境致子代糖尿病发病易感性增加的可能机制，为源头防控胎儿源性糖尿病发病、促进糖尿病防治前移提供理论依据。

糖尿病发病机制复杂，从妊娠期糖尿病的宫内环境暴露研究胎儿源性糖尿病的发病机制是该研究领域的崭新重大课题。本课题组通过利用GDM小鼠模型，明确了GDM宫内糖尿病环境可上调子代胰岛素通路蛋白FoxO1的表达，同时伴随着甲基化转移酶DNMT3A的上调，印记基因IGF2/H19的下调，与GDM子代胰岛素抵抗易感性相关。通过chip-qPCR及体外干扰实验进一步明确了FoxO1与DNMT3A的相互作用及调控关系，并通过体外胰岛素刺激实验进一步证明了GDM子代出现胰岛素抵抗归因于宫内高胰岛素环境，从而揭示了GDM宫内高胰岛素环境致子代胰岛素抵抗易感性增加的可能机制，为源头防控胎儿源性糖尿病发病、促进糖耐哦病防治前移提供理论依据。