Pulmonary epithelial cell differentiation in the nitrofen-induced congenital diaphragmatic hernia.

BACKGROUND/AIM: In congenital diaphragmatic hernia (CDH), there is pulmonary neuroendocrine cell (PNEC) hyperplasia and Clara (nonendocrine) cell hypoplasia, the meaning of which remains unknown. In embryonic/fetal lung, an intricate cross talk between Notch pathway and basic helix-loop-helix transcription factors Mash1 and Hes1 determines the balance between endocrine and nonendocrine epithelial cell fate. Differences at the molecular level in pulmonary epithelial cell differentiation between control and CDH hypoplastic lungs were investigated.

MATERIAL AND METHODS

The nitrofen-induced CDH rat model was used. At 15.5 days postconception (dpc), fetuses were assigned to 2 experimental groups: control and nitrofen (exposed to nitrofen, without CDH), whereas at 17.5, 19.5, and 21.5 dpc, fetuses were assigned to 3 experimental groups: control, nitrofen, and CDH (exposed to nitrofen, with CDH). The fetal lungs were processed for expression quantification of CC10, Hes1, Mash1, and Dll1 by real-time polymerase chain reaction.

RESULTS

In control fetuses, expression of all studied genes increased with gestational age. In nitrofen-exposed fetal lungs, endocrine cell marker Mash1 was downregulated only at the earliest studied gestational age, whereas Dll1 expression levels were significantly increased in the CDH group at 19.5 and 21.5 dpc. Regarding nonendocrine markers, Hes1 presented increased expression at 15.5 and 19.5 dpc, whereas CC10 was downregulated at 17.5 and 19.5 dpc but not at term.

CONCLUSIONS

This study suggests that PNEC hyperplasia in CDH fetal lung is likely because of Notch signaling deregulation, whereas Clara cell hypoplasia in CDH lungs could be a consequence of protein synthesis delay, reflecting a functional maturation hindrance and not a cell fate commitment problem.