Insulin receptor is downregulated in the nitrofen-induced hypoplastic lung.
OBJECTIVE
The pathogenesis of pulmonary hypoplasia in congenital diaphragmatic hernia (CDH) is still poorly understood. During fetal lung development, the insulin receptor (IR) plays an important role by mediating the cellular uptake of glucose, which is a major substrate for the biosynthesis of surfactant phospholipids. In fetal rat lung, IR gene expression has been revealed on type II pneumocytes. Recent studies have demonstrated that downregulation of pulmonary IR in late gestation causes pulmonary hypoplasia by inhibition of surfactant synthesis. We hypothesized that pulmonary gene expression of IR is downregulated during the late stages of lung development in the nitrofen-induced CDH model.


METHODS
Timed pregnant Sprague-Dawley rats were exposed to either olive oil or nitrofen on day 9 of gestation (D9). Cesarean deliveries were performed on D15, D18, and D21. Fetal lungs were divided into 3 groups: control, nitrofen without CDH (CDH[-]), and nitrofen with CDH (CDH[+]) (n = 8 at each time-point, respectively). Relative messenger RNA (mRNA) levels of IR were determined by using real time reverse transcription polymerase chain reaction. Immunohistochemistry was performed to evaluate protein expression of IR.


RESULTS
Relative expression levels of IR mRNA on D21 were significantly decreased in CDH(-) and CDH(+) group (3.99 +/- 1.50 and 5.14 +/- 0.99, respectively) compared to control (7.45 +/- 3.95; P < .05). Immunohistochemistry showed decreased IR expression in the proximal alveolar epithelium on D21 in hypoplastic lungs compared to control lungs.


CONCLUSIONS
Downregulation of IR gene and protein expression in hypoplastic lung during late stages of lung development may interfere with normal surfactant synthesis, causing pulmonary hypoplasia in the nitrofen-induced CDH model.
Ruttenstock
Elke
E
Children's Research Center, Our Lady's Children's Hospital, Dublin 12, Ireland.
Doi
Takashi
T
Dingemann
Jens
J
Puri
Prem
P
D000818:Animals; D015536:Down-Regulation; D018507:Gene Expression Regulation, Developmental; D006548:Hernia, Diaphragmatic / Q000196:embryology* / Q000473:pathology; D065630:Hernias, Diaphragmatic, Congenital; D008168:Lung / Q000002:abnormalities* / Q000196:embryology* / Q000473:pathology; D010647:Phenyl Ethers; D051381:Rats; D017207:Rats, Sprague-Dawley; D011972:Receptor, Insulin / Q000235:genetics / Q000378:metabolism*
D010647:Phenyl Ethers; D011972:Receptor, Insulin; C007350:nitrofen