心脏发育相关的微小RNA差异表达分析  

Experimental analysis of microRNAs related to cardiac development differential expression

作　　者：施翔翔[1] 来丹丹[1] 章佳颖[1] 褚茂平[1] 王赛斌[1] 倪秋明[1] 黄晓燕[1] 张怀勤[1] 杨德业[1] 

机构地区：[1]温州医学院附属第一医院心血管内科温州医学院心血管生物和基因研究所,325000

出　　处：《中华医学杂志》2009年第20期1416-1420,共5页National Medical Journal of China

基　　金：基金项目：国家自然科学基金（30571050）;浙江省温州市科技发展计划资助项目（Y2006A017）

摘　　要：目的探讨微小RNA（miRNA）在心脏发育中的作用。方法建立先天性心脏病-室间隔缺损相关基因Pax-8敲除小鼠模型Pax·8KO-／-（纯合子）及Pax-8KO＋／-（杂合子），提取纯合子和杂合子小鼠心脏总RNA，采用含有567种miRNA的芯片进行检测并分析miRNA表达谱的差异，并通过荧光实时定量PCR技术对结果进行验证。结果纯合子小鼠比杂合子小鼠左心室腔扩大，左心室壁肥厚，室间隔增厚，左室乳头肌肥大，心脏呈球形。超声结果显示杂合子小鼠左室收缩末期内径[（LVIDs）（1．23±0．25）mm比（1．93±0．50）mm]和左室舒张末期内径[（LVIDd）（2．13±0．18）mm比（2．89±0．29）]均明显小于纯合子小鼠，差异均有统计学意义（均P〈0．01）心肌左心室壁以及室间隔组织可见大量的凋亡心肌细胞。在纯合子小鼠发现miRNA表达谱中差异表达的miRNA共10个，其中2个miRNA表达下调，8个miRNA表达上调。荧光实时定量RT-PCR结果与芯片结果基本符合。结论miRNA参与心脏发育过程，在先天性心脏病室间隔缺损的发病机制中可能发挥重要作用。Objective To investigate the differential expression profiles of microRNAs in the knockout Pax-8 mice by miRNA microarray analysis and study the function of microRNA during cardiac development. Methods The knockout Pax-8 mice model was established and the total RNA derived from Pax-8 KO -/- and Pax-8 KO ＋/- mice heart. MicroRNA microarray containing 567 mammalian microRNA probes was used to investigate the microRNAs differential expression between Pax-8 KO -/- and Pax-8 KO ＋/- mice. The candidates of microRNAs were confirmed by real time RT-PCR assay. Results The heart of Pax-8 KO - / - mice became spheroidal. Left ventricle enlargement, left ventricular wall and interventricular septum thickening and papillary muscles in left ventricle enlargement were found. Furthermore, many apoptotic cells were discovered in left ventricular wall and interventricular septum in Pax-8 KO -/- mice. The MicroRNA microarray result displayed 10 microRNAs differential expressions, in which 2 microRNAs became down-regulated and 8 microRNAs up-regulated by more than two folds in Pax-8 KO - / - mice. This was in accordance with the result of real-time RT-PCR. Conclusion Some microRNAs may play important roles in cardiac development and ventricular septal defect pathogenesis.

关 键 词：室间隔缺损 小鼠 基因敲除 微RNAS 

分 类 号：R686[医药卫生—骨科学]