Pax8基因在心脏发育中蛋白表达下调.docVIP

Pax-8基因在心脏发育中蛋白表达下调 　　作者：倪秋明，章佳颖，来丹丹，褚茂平，王赛斌，黄晓燕，张怀勤，杨德业 作者单位：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，浙江 温州 325000 　　【摘要】 目的:观察Pax-8基因在ALK3基因敲除小鼠心脏中的蛋白表达情况，探讨Pax-8基因在心脏发育中的作用。方法:基因芯片(microarray)和定量逆转录-聚合酶链反应测定ALK3基因的下游基因，免疫组化法检测13.5 d小鼠胚胎心脏特异性ALK3基因敲除小鼠胚胎中Pax-8蛋白的表达，连续病理切片及HE染色观察刚出生的Pax-8基因敲除小鼠的心脏形态。结果：在心脏特异的ALK3基因敲除小鼠中，转录因子Pax-8基因mRNA的表达水平上被下调7.1倍。免疫组化结果显示：13.5 d小鼠胚胎心脏特异性 ALK3基因敲除的纯合子小鼠Pax-8基因在蛋白水平与杂合子相比也明显下调。Pax-8基因敲除小鼠纯合子有明显的室间隔和左心室肥厚，心脏呈球形。结论:Pax-8基因是ALK3基因的下游基因，与心脏发育有密切关系。 　　【关键词】 Pax-8基因;室间隔缺损;免疫组化;蛋白表达 　　Down-regulated protein expression level of Pax-8 gene during heart development in mice NI Qiu-ming，ZHANG Jia-ying，LAI Dan-dan，CHU Mao-ping，WANG Sai-bin，HUANG Xiao-yan，ZHANG Huai-qin，YANG De-ye. Department of Cardiology， the First Affiliated Hospital Institute of Cardiovascular Biology Gene， Wenzhou Medical College， Wenzhou，325000 　　Abstract: Objective: To investigate the level of Pax-8 protein expression in the condi-tional ALK3 kockout mice heart and study the role of Pax-8 gene in the cardiac development. Methods: The microarray and real time quantitative RT-PCR were used to investigate the down-stream genes of ALK3. Immunohistochemical technology was used to detect the level of Pax-8 protein expression in the 13.5 d heart derived from a-MHC Cre+/-，ALK3 F/- and a-MHC Cre+/-，ALK3 F/+ mice. To study the contribution of Pax-8 gene in the cardiac development，the heart derived from pax-8 kockout mice was overviewed using consecutive pathological section and HE staining method. Results:The level of Pax-8 mRNA expression was down-regulated 7.1 fold in the embryonic heart (11.5 days) of a-MHC Cre+/-，ALK3 F/-. The level of Pax-8 protein expression was also remarkably down-regulated in the embryonic heart (13.5 days) of a-MHC Cre+/-，ALK3 F/-. The heart of pax-8 KO-/-mice became spheroidal，the left ventricle enlarged， left ventricular wall and interventricular septum thickened， papillary muscles in the left ventricle enlarged. Conclusion: Pax-8 gene is