大鼠胚胎胰腺发育后期基因表达谱的研究  

Embryonic pancreatic gene expression profiles in rats at developing anaphase

作　　者：袁庆新[1] 仲燕[2] 刘超[1] 刘翠萍[1] 袁栎[2] 周锦勇[2] 滕丽萍[2] 胡静静[2] 德伟[2] 

机构地区：[1]南京医科大学第一附属医院内分泌科,江苏省南京市210029 [2]南京医科大学生化教研室,江苏省南京市210029

出　　处：《中国临床康复》2005年第39期81-83,i0002,共4页Chinese Journal of Clinical Rehabilitation

基　　金：江苏省科委应用基础资助项目(BK2001119)~~

摘　　要：目的:研究大鼠胚胎发育至第15.5和18.5天胰腺基因表达谱,探讨特异性基因表达在正常内外分泌胰腺发育及糖尿病中所起的作用。方法:①实验于2003-09/2004-06在南京医科大学生化与分子生物学实验室完成。选用SD大鼠,雌雄各10只,将雌雄鼠按1∶2合笼,分别于胚胎发育至第15.5和18.5天,剖腹取出孕子宫,分离胚胎。②应用显微分离及提取技术获得胚胎发育不同阶段胰腺组织并提取RNA,采用高密度寡核苷酸芯片(Affemetrix芯片)对胚胎发育至第15.5天和第18.5胚胎胰腺进行基因转录水平分析,用生物信息学方法分析具体基因的表达情况。结果:胚胎发育至第18.5天与胚胎发育至第15.5天相比,差异表达2倍以上的基因共1319条。其中表达上升的基因664条,表达下降的基因655条,功能已知基因占63%,表达序列标识占37%。控制胰腺细胞分化的转录因子明显下调,与功能代谢相关的转录因子有上升趋势,内外分泌酶类、激素类表达有上升趋势,骨架蛋白有下降趋势,与激素、酶类分泌相关的因子基本无表达。结论:大鼠胚胎胰腺发育至第15.5～18.5天,内分泌典型胰岛结构及外分泌腺泡、导管结构已基本形成,功能细胞分化基因表达下降,而细胞代谢调节功能基因表达上升,因此,在大鼠胚胎发育至第15.5天胰腺,功能细胞的代谢调节功能开始完善。AIM： To investigate pancreatic gene expression profiles and the effect of some specific genes on normal endocrine/exoerine pancreatic development and diabetes during embryonic clay 15.5 and embryonic clay 18.5 in rats. METHODS： ①The experiment was conducted at the Laboratory of Biochemistry and Molecular Biology, Nanjing Medical University from September 2003 to June 2004. Ten male and female （half and halo SD rats were selected. The female and male rats were put into cages according to the proportion of 1 to 2. Pregnant womb was gained by cutting the belly open to separate embryo at the embryonic clay 15.5 and embryonic clay 18.5, respectively. ② Embryonic panereata at different phase were isolated under microscope and extracting technique and the RNA was extracted. Gene transcription was analyzed at the embryonic clay 15.5 and embryonic clay 18.5 with high-density oligonueleotide core （Affemetrix core）. Its gene expression profiles were tested using bioinformaties method. RESULTS： Compared the embryonic clay 18.5 and embryonic clay 15.5, 1 319 genes were expressed differentially 2-fold above, in which 664 genes were up-regulated, and 655 genes were down-regulated, including 63% definite genes and 37% expressed rank sign. The transcription factor that controlled pancreatic cell differentiation reduced significantly, and those having relation with functional metabolism increased. Endocrine/exocrine enzyme and hormone enhanced; skeleton protein decreased; factors related with excretion of hormone and enzyme had no expression mostly. CONCLUSION： During embryonic clay 15.5 and embryonic clay 18.5 in rats, typical pancreatic islets structure of endocrine with exocrine aeinus and canal structure has formed generally. Expression of differentiation gene of functional cells reduces, while expression of metabolic regulation gene increases. Therefore, metabolic regulation function is improving at the embryonic day 15.5.

关 键 词：胰腺 胎儿发育 寡核苷酸序列分析 基因表达 

分 类 号：R329.449[医药卫生—人体解剖和组织胚胎学]