【摘要】 目的:寻找先天性心脏病室间隔缺损相关基因——转录因子Pax-8保护细胞凋亡的途径。方法:提取Pax-8基因敲除小鼠纯合子(Pax-8 KO-/-)和杂合子(Pax-8 KO+/-)的心脏总RNA,利用小鼠基因的基因芯片检测两组小鼠基因表达水平,找出差异表达的基因,并经半定量RT-PCR和荧光实时定量PCR技术初步筛选出转录因子Pax-8的下游基因。结果:基因芯片检测发现,与Pax-8 KO+/-组相比,在Pax-8 KO-/-小鼠中有25个基因表达下调,17个基因表达上调。用半定量RT-PCR验证发现,nuclear receptor sub-family4,group A,member 1(NR4A1)基因在Pax-8 KO-/-组上调。定量RT-PCR亦证实在Pax-8 KO-/-组NR4A1基因的表达水平较Pax-8 KO+/-组及Pax-8+/+(野生型)组分别上调1.53倍和4.79倍(P<0.01)。结论:NR4A1基因受Pax-8基因调控,在Pax-8保护细胞凋亡途径中发挥作用。更多还原

【Abstract】 Objective: To find the possible genic pathway for the anti-apoptosis function of the ventricular septal defeet related to Pax-8.Method: The total RNA derived from the heart of Pax-8 KO-/-and Pax-8 KO+/-mice was extracted.Mouse genome DNA microarray was used to detect the differential expression level between the Pax-8 KO-/-and the Pax-8 KO+/-mice heart.The candidate genes were confirmed by RT-PCR and real time RT-PCR assay.Results: Microarray results showed that 25 genes were down-regulated and 17 were up-regulated in the Pax-8 KO-/-mice compared to the Pax-8 KO+/-mice.Nuclear receptor subfamily 4,group A,member 1(NR4A1)was proved to be up-regulated by RT-PCR in the Pax-8 KO-/-mice heart.Real time RT-PCR results revealed that NR4A1 in the Pax-8 KO-/-mice was 1.53 and 4.79 fold as much as in the Pax-8 KO+/-and the Pax-8+/+ mice(P <0.01).Conclusion: The NR4A1 gene is one of the downstream genes of the Pax-8 and probably evolved and played an important role in the mechanism of ventricular septum defect.更多还原