title: Expression of hepatic lipid droplets is decreased in the nitrofen model of congenital diaphragmatic hernia.
abstract: BACKGROUND
  Prenatal mortality in newborn infants with congenital diaphragmatic hernia (CDH) has been attributed to increased amounts of liver hernia ion through the diaphragmatic defect. Antenatal studies in human and rodent fetus with CDH further demonstrated a contribution of the developing liver in the pathogenesis of CDH. The abnormal hepatic growth in experimental animal models, therefore, indicates a disruption of normal liver development in CDH. However, the underlying structural, histological and functional changes in the liver of animals with CDH remain unclear. We design this study to test the hypothesis that the morphological and cellular liver development is altered in the nitrogen-induced CDH model.
  
  
  METHODS
  Pregnant rats were exposed to either olive oil or nitrofen on day 9 of gestation (D9). Livers and chest were harvested on D21 and divided into two groups: control (n = 8), nitrofen with CDH (CDH, n = 8). Haematoxylin-eosin (Straub et al. Histopathology 68:617-631, 2013) staining was performed to evaluate underlying morphological changes. Apoptosis was checked by using TUNEL staining and apoptotic cell number was counted on 16-16 slides in 25 fields by two independent viewers. Hepatic lipid droplet expressions were evaluated by hepatic adipose differentiation-related protein (ARDP) expression.
  
  
  RESULTS
  Compared to controls markedly increased hypertrophy was seen in CDH group. Significantly increased apoptotic cell numbers were detected in CDH group compared to controls (5.1 ± 1.5 vs 2.1 ± 0.6) (p < 0.05). The relative mRNA expression levels of ARDP were significantly reduced in CDH group compared to controls. Immunohistochemistry showed markedly decreased hepatic ADRP immunoreactivity in CDH fetuses compared to controls.
  
  
  CONCLUSIONS
  Our findings provide strong evidence of hepatic hypertrophy and increased cell apoptosis in the liver of nitrofen-induced CDH. These morphological changes may affect liver lipid droplet expression function.
authors.affiliation: Our Lady's Children's Hospital, National Children's Research Centre, Dublin 12, Ireland.
authors.forename: Hiromizu
authors.initials: H
authors.lastname: Takahashi
authors.affiliation: Our Lady's Children's Hospital, National Children's Research Centre, Dublin 12, Ireland.
authors.forename: Balazs
authors.initials: B
authors.lastname: Kutasy
authors.affiliation: Our Lady's Children's Hospital, National Children's Research Centre, Dublin 12, Ireland.
authors.forename: Florian
authors.initials: F
authors.lastname: Friedmacher
authors.affiliation: Our Lady's Children's Hospital, National Children's Research Centre, Dublin 12, Ireland.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Takahashi
authors.affiliation: Our Lady's Children's Hospital, National Children's Research Centre, Dublin 12, Ireland. prem.puri@ncrc.ie.
authors.forename: Prem
authors.initials: P
authors.lastname: Puri
mesh_terms: D000818:Animals; D017209:Apoptosis; D004195:Disease Models, Animal; D065630:Hernias, Diaphragmatic, Congenital / Q000378:metabolism*; D066292:Lipid Droplets / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D010647:Phenyl Ethers; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D010647:Phenyl Ethers; C007350:nitrofen
keywords: ARDP; CDH; Congenital diaphragmatic hernia; Liver; Malnutrition; Nitrofen