2型糖尿病性心脏病变早期相关基因的筛选和分析

Screening and analysis of early cardiopathology-related gene in type 2 diabetes mellitus

目的 筛选 2型糖尿病性心脏病变早期相关基因并探讨其发病机制。方法 (1)SD大鼠高热量饮食 2个月后链脲佐菌素 (STZ ,15mg/kg)一次性尾静脉注射 ,建立 2型糖尿病大鼠模型。(2 )分别于模型建立后 0 5、1 5、6 0个月取大鼠心脏组织进行电镜观察。 (3)模型建立后 15d抽提大鼠心脏组织总RNA进行荧光标记的mRNA差异显示分析 ,用Northernblot证实差异表达基因并对未知序列进行生物信息学分析。结果 (1)建立的 2型糖尿病模型大鼠 ,具有外周胰岛素抵抗和胰岛功能仅轻微受损等特征。 (2 )心肌超微结构观察结果证实糖尿病心脏病变出现于模型建立后 1 5个月的大鼠心脏 ,并随着病程延长而加重。 (3)应用mRNA差异显示分析技术筛选得到的克隆中 ,已知序列有肌型肉碱棕榈酰转移酶 ;未知序列分别命名为DCM1、2、5、6、8、11、12、16 ,已提交Genbank。结论 糖尿病心脏病变早期在超微结构发生改变前已出现能量代谢途径相关酶类基因表达差异 ,对其进一步研究将为早期干预逆转糖尿病心脏病变提供思路。

Objective To screen and analyze early cardiopathology related gene in type 2 diabetes mellitus (DM) Methods (1) To develop a rat model of type 2 DM, SD rats were injected with streptozotocin(STZ, 15 mg/kg)after high caloric diet had been given for two months (2)After establishment of the model for half a month, one and a half month and six months respectively, dissected myocardium was observed under electron microscope (3)After establishment of the model for half a month, myocardium was analyzed with fluorescence labelled mRNA differential display Positive clones were BLAST after conformed by Northern blot Via the application of bioinformatics, the function and structure of unknown sequences were predicted Results (1) The model of type 2 DM developed with the character of lightly impaired islet function and insulin resistance (2) Diabetic cardiomyopathy was verified by the ultrastructural change of myocardium which became serious with time (3)Screened and conformed clones were muscle carnitine palmitoyltransferase 1 and 8 and novel sequences were named DCM1?2?5?6?8?11?12?16, which had been submitted to Genbank DCM 5 was predicted to play a role of catalysis or signal conduction via the application of bioinformatics Conclusions The altered mRNA expression of these genes suggested that they were candidates for a role in the development of cardiopathy prior to the ultrastructural alterations of myocardium The change of key enzyme in energy metabolism may be the initial factor during heart functional disorder Further investigation was required for reverse intervention of diabetic cardiopathy in early period