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

血管内皮生长因子治疗胎儿先天性膈疝的效果。

• 批准号: 14571699
• 负责人: KOUCHI Katsunor
• 金额: $ 2.3万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571699/
• 关键词: congenital diaphragmatic hernia; fetal therapy; vascular endothelial growth factor; VEGF

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

目的：最近的研究报告称，经父母诊断患有先天性膈疝 (CDH) 的新生儿预后较差。父母诊断的先天性膈疝新生儿肺动脉肌层厚度。这些机制在出生后不久就引起了严重的肺动脉高压。肺动脉肌层厚度的机制尚不清楚。为了揭示这一机制，我们分析了血管重塑的主要因素； CDH模型大鼠血管内皮生长因子(VEGF)、血管生成素(Ang)。方法：用硝基芬100mg溶于1ml橄榄油中，给孕9.5天的孕鼠口服，制成CDH模型大鼠。在 GA 17、19、21 天时，通过剖腹产分娩胎儿。在显微镜下获得肺组织。我们将胎儿分为两组；显微镜下未观察到CDH-；CDH，显微镜下未观察到CDH+；CDH。对照组是在 GA 9.5 天的怀孕大鼠中口服 1ml 橄榄油。我们分析了 VEGF（其受体）的 mRNA 水平； FLK1、Ang-1、Ang-2、其受体； TIE2、PDGFb及其受体；通过RT-PCR检测PDGFbr。 结果：与CDH-GA21相比，CDH+GA 21中VEGF的mRNA水平显着增加。两组无显着性差异； CDH+ GA21 和对照 GA21 的 VEGF mRNA 水平。三组中FLK1 mRNA水平无差异。相反，与CDH-和对照GA21相比，CDH+GA 21中Ang-1的mRNA水平显着降低。 Ang-2 mRNA表现出广泛的表达水平，各组之间未观察到相关性。 TIE2 mRNA水平在三组中没有差异。 PDGFb和PDGFbr在三组和各GA中均无差异。讨论：在CDH大鼠胎儿中，VEGF和Ang-1 mRNA在CDH+GA21中表现出不同的表达水平。这表明CDH在胎儿期发生了血管重塑。这些重塑因子的变化被认为引起了肺动脉肌层的退化。然而，患有 CDH 的新生儿显示出肺动脉肌层的显着厚度。 PDGFb和PDGFbr是血管平滑肌增殖的主要因素。在我们的研究中，这些 mRNA 水平在 CDH+ 组和对照组中没有显示出差异。根据这些结果，其他基因可能会诱导 CDH 中平滑肌细胞的过度增殖。较少的