CAJ | 학술논문

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-8 KO-/-(纯合子)及Pax-8 KO +/-(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.
목적 탐토미소RNA(miRNA)재심장발육중적작용.방법 건립선천성심장병-실간격결손상관기인Pax-8고제소서모형Pax-8 KO-/-(순합자)급Pax-8 KO +/-(잡합자),제취순합자화잡합자소서심장총RNA,채용함유567충miRNA적심편진행검측병분석miRNA표체보적차이,병통과형광실시정량PCR기술대결과진행험증.결과 순합자소서비잡합자소서좌심실강확대,좌심실벽비후,실간격증후,좌실유두기비대,심장정구형.초성결과현시잡합자소서좌실수축말기내경(LVIDs)[(1.23±0.25)mm비(1.93±0.50)mm]화좌실서장말기내경(LVIDd)[(2.13±0.18)mm 비(2.89±0.29)]균명현소우순합자소서,차이유통계학의의(균P<0.01)심기좌심실벽이급실간격조직가견대량적조망심기세포.재순합자소서발현miRNA표체보중차이표체적miRNA공10개, 기중2개miRNA표체하조,8개miRNA표체상조.형광실시정량RT-PCR결과여심편결과기본부합.결론 miRNA삼여심장발육과정,재선천성심장병실간격결손적발병궤제중가능발휘중요작용.
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.