摘要
目的研究心脏发育相关基因心肌转录因子NKX2.5和锌指转录因子GATA4在先天性心脏病(CHD)的相互作用关系。方法构建基因NKX2.5和GATA4真核表达质粒及脑钠肽启动子荧光素酶报告基因表达质粒,真核表达质粒经转染后,利用免疫共沉淀技术证明基因NKX2.5和GATA4两者是否有相互作用,通过荧光素酶分析技术分析基因NKX2.5和GATA4对脑钠肽启动子序是否存在协同激活作用。结果 p CMV-HA-NKX2.5与Pre-immune作用后蛋白表达量0,与Anti-Myc作用后蛋白表达量为3.63±1.23,与Input作用后蛋白表达量为2.95±1.05,NKX2.5可将GATA4沉淀下来;p CMV-Myc-GATA4与Pre-immune作用后蛋白表达量0,与Anti-Myc作用后蛋白表达量为5.63±2.06,与Input作用后蛋白表达量为5.75±2.86,GATA4可将NKX2.5沉淀下来;脑钠肽(BNP)启动子荧光表达质粒与两者共转染比分别单独转染NKX2.5和GATA4对脑钠肽启动子的激活作用更强。结论 NKX2.5和GATA4之间具有相互作用,在心脏发育过程中起着重要的协同作用。
Objective To investigate the interaction of cardiac transcription factors of cardiac developmental gene TN domain and the NK2-specific domain（ NKX2. 5） and zinc finger transcription factor GATA binding factor 4（ GATA4） in patients with congenital heart disease（ CHD）. Methods NKX2. 5 and GATA4 eukaryotic gene expression plasmid and brain natriuretic peptide promoter luciferase reporter gene expression plasmid were constructed. After a eukaryotic expression vector were transfected, the interactions of gene NKX2. 5 and GATA4 were proved by co-immunoprecipitation. And the synergistic activations of NKX2. 5 and GATA4 with brain natriuretic peptide promoter sequence were analyzed by fluorescein enzyme technology. Results Protein expression of p CMV-HA-NKX2. 5 with Pre-immune effects was 0,and protein expression after Anti-Myc was 3. 63 ± 1. 23,and the role of protein expression after Input was 2. 95 ± 1. 05,NKX2. 5 can GATA4 precipitate. Protein expression was 0 after p CMV-Myc-GATA4 and Pre-immune effects, and protein expression after Anti-Myc was5. 63 ± 2. 06, and the role of protein expression after Input was 5. 75 ± 2. 86,GATA4 can settle NKX2. 5 down. Promoter fluorescence expression of brain natriuretic peptide（ BNP）and the two co-transfected plasmids than separately transfected NKX2. 5 and GATA4 for BNP promoter activation stronger. Conclusion NKX2. 5 had an interaction with GATA4,which was an important synergy in the heart development process.
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2017年第18期1773-1775,共3页
The Chinese Journal of Clinical Pharmacology
基金
广东省医学科研基金面上基金资助项目(B2015134)
