Oue T, Yoneda A, Shima H, Taira Y, Puri P
Children's Research Centre, Our Lady's Hospital for Sick Children, Crumlin, Dublin 12, Ireland.
Pediatr Surg Int. 2002 May;18(4):221-6. doi: 10.1007/s003830100625.
Persistent pulmonary hypertension (PPH) in congenital diaphragmatic hernia (CDH) lung has been shown to be associated with structural changes in the pulmonary vasculature, including medial and adventitial thickening. Vascular endothelial growth factor (VEGF) is a potent mitogenic and permeability factor targeting predominantly endothelial cells. mRNA encoding VEGF is detected in all fetal tissues and is most abundant in fetal lung, kidney, and liver. Recently, antenatal dexamethasone (Dex) treatment has been shown to prevent pulmonary-artery structural changes in experimentally-produced CDH. The aim of this study was to investigate mRNA and protein levels of VEGF in CDH lung and to determine whether antenatal Dex treatment has any effect on the production of VEGF. A CDH model was induced in pregnant rats following administration of 100 mg nitrofen on days 9.5 of gestation (term=22 days). Dex 0.25 mg/kg was given on day 18.5 and 19.5. Cesarean section was performed on day 21 of gestation. The fetuses were divided into three groups: normal controls (NC, n=8); nitrofen-induced CDH (CDH, n=8); and nitrofen-induced CDH with antenatal Dex treatment (CDH-Dex, n=8). Protein and mRNA were extracted from the whole lung. VEGF protein was measured by ELISA assay and mRNA expression was evaluated by reverse transcription-polymerase chain reaction. Immunohistochemistry using anti-rat VEGF antibody was also performed in each group. VEGF protein as well as mRNA expression were significantly increased in the CDH group compared to the NC group, which was not affected by antenatal Dex treatment. VEGF immunoreactivity in pulmonary vessel walls was increased in the CDH and CDH-Dex groups compared to the NC group. The enhanced VEGF protein and mRNA expression in CDH lung suggests that increased local synthesis of VEGF may be responsible for the structural changes in the pulmonary vasculature in CDH lung. VEGF expression in CDH lung is not downregulated by antenatal Dex treatment.
先天性膈疝（CDH）患儿肺部的持续性肺动脉高压（PPH）已被证实与肺血管结构改变有关，包括中膜和外膜增厚。血管内皮生长因子（VEGF）是一种主要作用于内皮细胞的强效促有丝分裂和通透性因子。编码VEGF的mRNA在所有胎儿组织中均可检测到，在胎儿肺、肾和肝脏中含量最为丰富。最近的研究表明，产前地塞米松（Dex）治疗可预防实验性CDH中肺动脉结构的改变。本研究的目的是调查CDH患儿肺部VEGF的mRNA和蛋白水平，并确定产前Dex治疗是否对VEGF的产生有任何影响。在妊娠第9.5天（足月为22天）给怀孕大鼠注射100 mg硝呋烯腙，诱导建立CDH模型。在第18.5天和19.5天给予0.25 mg/kg Dex。在妊娠第21天进行剖宫产。将胎儿分为三组：正常对照组（NC，n = 8）；硝呋烯腙诱导的CDH组（CDH，n = 8）；以及产前接受Dex治疗的硝呋烯腙诱导的CDH组（CDH-Dex，n = 8）。从整个肺组织中提取蛋白和mRNA。通过ELISA法检测VEGF蛋白，通过逆转录-聚合酶链反应评估mRNA表达。每组还进行了使用抗大鼠VEGF抗体的免疫组织化学检测。与NC组相比，CDH组中VEGF蛋白以及mRNA表达均显著增加，而产前Dex治疗对此无影响。与NC组相比，CDH组和CDH-Dex组肺血管壁中的VEGF免疫反应性增加。CDH患儿肺部VEGF蛋白和mRNA表达增强表明，VEGF局部合成增加可能是CDH患儿肺部肺血管结构改变的原因。产前Dex治疗并未下调CDH患儿肺部的VEGF表达。
