Pereira-Terra Patrícia, Kholdebarin Ramin, Higgins Meghan, Iwasiow Barbara M, Correia-Pinto Jorge, Keijzer Richard
Division of Pediatric Surgery, Departments of Surgery, Pediatrics and Child Health and Physiology and Pathophysiology (Adjunct), University of Manitoba and Children's Hospital Research Institute of Manitoba, Biology of Breathing Theme, AE402-840 Sherbrook Street, Winnipeg, MB, R3A 1R9, Canada.
Pediatr Surg Int. 2015 Jul;31(7):659-63. doi: 10.1007/s00383-015-3703-2. Epub 2015 Apr 11.
Congenital diaphragmatic hernia (CDH) is characterized by a developmental defect in the diaphragm, pulmonary hypoplasia and pulmonary hypertension. NPAS3 is a PAS domain transcription factor regulating Drosophila tracheogenesis. NPAS3 null mice develop pulmonary hypoplasia in utero and die after birth due to respiratory failure. We aimed to evaluate NPAS3 expression during normal and abnormal lung development due to CDH.
CDH was induced by administering 100 mg/ml nitrofen to time-pregnant dams on embryonic day (E) 9 of gestation. Lungs were isolated on E15, E18 and E21 and NPAS3 localization was determined by immunohistochemistry and quantified using Western blotting.
We found that only E21 hypoplastic CDH lungs have reduced expression of NPAS3 in the terminal saccules. Western blotting confirmed the down-regulation of NPAS3 protein in the nitrofen-induced hypoplastic lungs.
We demonstrate for the first time that nitrofen-induced hypoplastic CDH lungs have reduced NPAS3 expression in the terminal saccules during the later stages of abnormal lung development. Our findings suggest that NPAS3 is associated with pulmonary hypoplasia in CDH.
先天性膈疝（CDH）的特征是膈肌发育缺陷、肺发育不全和肺动脉高压。NPAS3是一种调节果蝇气管生成的PAS结构域转录因子。NPAS3基因敲除小鼠在子宫内发生肺发育不全，并在出生后因呼吸衰竭死亡。我们旨在评估CDH导致的正常和异常肺发育过程中NPAS3的表达情况。
在妊娠第9天（E9）给怀孕的母鼠腹腔注射100mg/ml的硝呋烯腙以诱导CDH。在E15、E18和E21分离肺组织，通过免疫组织化学确定NPAS3的定位，并使用蛋白质免疫印迹法进行定量分析。
我们发现只有E21期发育不全的CDH肺组织终末囊泡中NPAS3表达降低。蛋白质免疫印迹法证实硝呋烯腙诱导的发育不全肺组织中NPAS3蛋白表达下调。
我们首次证明，在异常肺发育后期，硝呋烯腙诱导的发育不全CDH肺组织终末囊泡中NPAS3表达降低。我们的研究结果表明，NPAS3与CDH中的肺发育不全有关。
