The effect of vascular endothelial growth factor for the treatment of congenital diaphragmatic hernia in the fetus.

目的：近年来的研究报道，经父母诊断的先天性腹股沟疝（CDH）的新生儿预后不良。经肠外诊断的先天性腹股沟疝患儿肺动脉肌层增厚。这些机制在出生后不久就引起了严重的肺动脉高压。肺动脉肌层增厚的机制尚不清楚。方法：在孕9.5天的CDH大鼠中，用1 ml橄榄油灌服硝苯地平100 mg，制备CDH模型。分别于GA 17、19、21天行剖宫产术。在显微镜下获得肺组织。我们将胎儿分为两组：CDH-;CDH在显微镜下未观察到和CDH+;CDH在显微镜下未观察到。对照组口服GA 9.5的橄榄油1 ml 关于我们 怀孕的老鼠RT-PCR检测CDH+ GA 21组VEGF及其受体、FLK 1、Ang-1、Ang-2及其受体、TIE 2、PDGFb及其受体、PDGFbr mRNA水平。CDH+ GA 21组与对照组相比，VEGF mRNA水平无显著性差异。FLK 1 mRNA水平在三组间无差异。与此相反，Ang-1的mRNA水平在CDH+ GA 21中显著低于CDH-和对照GA 21。Ang-2 mRNA在各组中均呈广泛表达，但无相关性。TIE 2 mRNA在三组间无差异。结论：在CDH胎鼠中，VEGF和Ang-1 mRNA在CDH+ GA 21中表达水平不同。提示CDH在胎儿期就发生了血管重构。这些重塑因子的改变可能是导致肺动脉肌层退行性变的原因。新生儿CDH肺动脉肌层增厚明显。PDGFb和PDGFbr是促进血管平滑肌增殖的主要因子。在我们的研究中，这些mRNA水平在CDH+和对照组中没有差异。根据这些结果，推测其他基因在CDH中诱导平滑肌细胞过度增殖。少

Purpose : Recent studies reported that neonates with congenital diaphragmatic hernias (CDH) diagnosed parentally showed poor prognosis. Neonates with congenital diaphragmatic hernias diagnosed parentally had the thickness of muscular layer of lung artery. The mechanisms caused severe pulmonary hypertension soon after birth. The mechanisms of the thickness of muscular layer of lung artery have been unknown. To reveal this mechanism, we analyzed major factor of vascular remodeling ; vascular endothelial growth factor (VEGF), angiopoietin (Ang) in the CDH model rats.Methods : CDH model rat was made by oral intake of Nitrofen 100mg in 1ml of olive oil at GA 9.5 days pregnant rats. At GA 17, 19, 21 days, fetuses were delivered by cesarean section. The lung tissue was obtained under microscopy. We classified fetus into two groups ; CDH-;CDH was not observed under microscopy and CDH+;CDH was not observed under microscopy. The control group was made by oral intake of 1ml of olive oil at GA 9.5 … More days pregnant rats. We analyzed mRNA levels of VEGF, its receptor ; FLK1, Ang-1, Ang-2, its receptor ; TIE2, PDGFb and its receptor ; PDGFbr by RT-PCR.Results : The mRNA level of VEGF was significantly increased in CDH+ GA 21 compared than CDH- GA21. No significant difference was observed in two group ; CDH+ GA21 and Control GA21 in mRNA level of VEGF. The mRNA level of FLK1 was no difference in three groups. In contrast, the mRNA level of Ang-1 was significantly decreased in CDH+ GA 21 compared than CDH- and control GA21. The mRNA of Ang-2 showed wide expression levels and no relation was not observed in each group. TIE2 mRNA level was no difference in three groups. Both of PDGFb and PDGFbr was no difference in three groups and in each GA.Discussion : In the fetus of CDH rat, the mRNA of VEGF and Ang-1 showed different expression level in CDH+ GA21. This indicated that the vascular remodeling occurred by CDH during fetal periods. These changes of the remodeling factors were though to induce the regression of muscular layer of lung artery. Though, neonates with CDH showed the remarkable thickness of muscular layer of lung artery. PDGFb and PDGFbr was major factor for proliferation of smooth muscle of vessels. In our study, these mRNA level showed no difference in CDH+ and control group. According to these results, the other genes were supposed to induce hyper-proliferation of smooth muscle cells in CDH. Less