Decreased expression of GATA4 in the diaphragm of nitrofen-induced congenital diaphragmatic hernia.
Decreased expression of GATA4 in the diaphragm of nitrofen-induced congenital diaphragmatic hernia.
复制标题
除草醚诱导的先天性膈疝膈肌中 GATA4 表达降低。
DOI：
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
P. Puri
中科院分区：
文献类型：
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作者：
J. Dingemann;T. Doi;J. Gosemann;E. Ruttenstock;Nana Nakazawa;P. Puri
背景
先天性横隔疝(CDH)导致横隔膜缺陷的分子机制尚不清楚。转录因子GATA4对于横隔膜的正常发育是必不可少的。最近，GATA4基因的突变被认为与人类和啮齿动物的CDH有关。我们假设，在硝芬CDH模型中，隔膜GATA4的表达下调。
方法
妊娠第9天(D9)给予硝苯芬酯或赋形剂。分别于D13、D18或D21处死胎儿。显微解剖胸腹膜皱褶(n=20)和胎儿横隔膜(n=40)，将其分为CDH组和对照组。提取RNA和蛋白质。实时荧光定量聚合酶链式反应检测GATA4基因表达水平。用酶联免疫吸附试验和免疫组织化学方法检测蛋白水平。
结果
CDH组在d13(mRNA15.96±6.99vs.38.10±5.01，p<；0.05)、d18(mRNA10.45±1.84vs.17.68±2.11，蛋白2.59±0.06vs.4.58±0.35p<；0.05)和d21(mRNA4.31±0.83 vs.6.87±0.88，蛋白0.16±0.08vs.1.26±0.49，p<；0.05)均显著低于对照组。在D13、D18和D21的CDH横隔部，GATA4的免疫反应活性显著降低。
结论
我们首次提供证据表明，在硝酸甘油诱导的CDH模型中，GATA4的横隔膜表达下调，提示GATA4的低表达可能损害了硝普芬诱导的CDH的横隔膜发育。
BACKGROUND
The molecular mechanisms underlying the diaphragmatic defect in congenital diaphragmatic hernia (CDH) are still poorly understood. The transcription factor GATA4 is essential for normal development of the diaphragm. Recently, mutations in the GATA4 gene have been linked to human and rodent CDH. We hypothesized that diaphragmatic GATA4 expression is downregulated in the nitrofen CDH model.
METHODS
Pregnant rats received Nitrofen or vehicle on day 9 of gestation (D9). Fetuses were sacrificed on D13, D18, or D21. Pleuroperitoneal folds (n=20) and fetal diaphragms (n=40) were (micro) dissected and divided into CDH group and controls. RNA and protein were extracted. GATA4 mRNA levels were determined by real-time PCR. Protein levels were determined by ELISA and Immunohistochemistry.
RESULTS
mRNA levels and Protein levels were significantly decreased in the CDH group compared to controls on D13 (mRNA 15.96±6.99 vs. 38.10±5.01, p<0.05), D18 (mRNA 10.45±1.84 vs. 17.68±2.11, Protein 2.59±0.06 vs. 4.58±0.35 p<0.05) and D21 (mRNA 4.31±0.83 vs. 6.87±0.88, Protein 0.16±0.08 vs. 1.26±0.49, p<0.05). Immunoreactivity of GATA4 was markedly decreased in CDH-diaphragms on D13, D18, and D21.
CONCLUSIONS
We provide evidence for the first time that diaphragmatic expression of GATA4 is downregulated in the nitrofen model, suggesting that decreased expression of GATA4 may impair diaphragmatic development in nitrofen-induced CDH.
影响因子：
10.5
作者：
Kuo, CT;Morrisey, EE;Leiden, JM
通讯作者：
Leiden, JM
影响因子：
2.7
作者：
Jay, Patrick Y.;Bielinska, Malgorzata;Wilson, David B.
通讯作者：
Wilson, David B.