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Pediatric Surgery International

, Volume 30, Issue 10, pp 1023–1029 | Cite as

Evidence for decreased lipofibroblast expression in hypoplastic rat lungs with congenital diaphragmatic hernia

  • Florian Friedmacher
  • Naho Fujiwara
  • Alejandro Daniel Hofmann
  • Hiromizu Takahashi
  • Jan-Hendrik Gosemann
  • Prem Puri
Original Article

Abstract

Purpose

Pulmonary hypoplasia (PH) is a serious condition in newborns with congenital diaphragmatic hernia (CDH). Lipid-containing interstitial fibroblasts (LIFs) play an essential role in fetal lung maturation by stimulating alveolarization and lipid homeostasis. In rodents, LIFs are first evident during the canalicular phase of lung development with a significant increase over the last 4 days of gestation. Adipocyte differentiation-related protein (ADRP), a functional lipogenic molecular marker characterizing LIFs, is highly expressed in fetal lungs during this critical time period. We hypothesized that LIF expression in hypoplastic rat lungs is decreased in the nitrofen-induced CDH model, which is accompanied by reduced alveolar ADRP expression and lipid content.

Methods

On embryonic day 9.5 (E9.5), time-mated rats received either nitrofen or vehicle. Fetuses were sacrificed on selected time points E18.5 and E21.5, and dissected lungs were divided into controls and CDH-associated PH. Pulmonary gene expression levels of ADRP were determined by quantitative real-time polymerase chain reaction. ADRP immunohistochemistry and oil red O staining were used to assess pulmonary protein expression and lipid content. Immunofluorescence double staining for alpha smooth muscle actin, which is known to be absent in LIFs, and lipid droplets was performed to evaluate the pulmonary expression of this specific subset of fibroblasts.

Results

Relative mRNA expression of ADRP was significantly reduced in lungs of CDH-associated PH on E18.5 and E21.5 compared to controls. ADRP immunoreactivity and lipid staining were markedly diminished in alveolar mesenchymal cells of CDH-associated PH on E18.5 and E21.5 compared to controls. Confocal laser scanning microscopy demonstrated markedly decreased LIF expression in alveolar interstitium of CDH-associated PH on E18.5 and E21.5 compared to controls.

Conclusion

Decreased pulmonary LIF expression during late gestation suggests impaired LIF functioning in the nitrofen-induced CDH model, which may cause disruption in fetal alveolarization and lipid homeostasis, and thus contribute to the development of PH.

Keywords

Lipofibroblast Adipocyte differentiation-related protein Pulmonary hypoplasia Congenital diaphragmatic hernia Nitrofen 

Notes

Acknowledgments

This research was financially supported by the National Children’s Research Centre and the Children’s Medical and Research Foundation.

Conflict of interest

The authors declare that this research was conducted in the absence of any commercial or financial relationship that could be construed as a potential conflict of interest.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Florian Friedmacher
    • 1
    • 2
  • Naho Fujiwara
    • 1
  • Alejandro Daniel Hofmann
    • 1
  • Hiromizu Takahashi
    • 1
  • Jan-Hendrik Gosemann
    • 1
  • Prem Puri
    • 1
    • 3
  1. 1.National Children’s Research CentreOur Lady’s Children’s HospitalDublin 12Ireland
  2. 2.Department of Pediatric and Adolescent SurgeryMedical University GrazGrazAustria
  3. 3.Conway Institute of Biomolecular and Biomedical Research, School of Medicine and Medical ScienceUniversity College DublinDublinIreland

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