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Increased vascular endothelial growth factor peptide and gene expression in hypoplastic lung in nitrofen induced
1Children's Research Centre, Our Lady's Hospital for Sick Children, Crumlin, Dublin 12, Ireland.
Abstract:
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.
