Skip to main content

Advertisement

SpringerLink
Log in

Expression of hepatic lipid droplets is decreased in the nitrofen model of congenital diaphragmatic hernia

  • Original Article
  • Published:
Pediatric Surgery International Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Colvin J, Bower C, Dickinson JE et al (2005) Outcomes of congenital diaphragmatic hernia: a population-based study in Western Australia. Pediatrics 116:e356–e363

    Article  PubMed  Google Scholar 

  2. Straub BK, Gyoengyoesi B, Koenig M et al (2013) Adipophilin/perilipin-2 as a lipid droplet-specific marker for metabolically active cells and diseases associated with metabolic dysregulation. Histopathology 62:617–631

    Article  PubMed  Google Scholar 

  3. Stege G, Fenton A, Jaffray B (2003) Nihilism in the 1990s: the true mortality of congenital diaphragmatic hernia. Pediatrics 112:532–535

    Article  PubMed  Google Scholar 

  4. Robinson PD, Fitzgerald DA (2007) Congenital diaphragmatic hernia. Paediatric respiratory reviews 8:323–334 (quiz 334–325)

    Article  PubMed  Google Scholar 

  5. Werner NL, Coughlin M, Kunisaki SM et al (2015) Prenatal and postnatal markers of severity in congenital diaphragmatic hernia have similar prognostic ability. Prenatal Diag. doi:10.1002/pd.4721

  6. Zamora IJ, Olutoye OO, Cass DL et al (2014) Prenatal MRI fetal lung volumes and percent liver herniation predict pulmonary morbidity in congenital diaphragmatic hernia (CDH). J Pediatr Surg 49:688–693

    Article  PubMed  Google Scholar 

  7. Nagata K, Usui N, Terui K et al (2015) Risk factors for the recurrence of the congenital diaphragmatic hernia-report from the long-term follow-up study of Japanese CDH study group. Eur J Pediatr Surg 25:9–14

    PubMed  Google Scholar 

  8. Terui K, Nagata K, Hayakawa M et al (2015) Growth assessment and the risk of growth retardation in congenital diaphragmatic hernia: a long-term follow-up study from the Japanese congenital diaphragmatic hernia study group. Eur J Pediatr Surg [Epub ahead of print]

  9. Langwieler T, Fiegel HC, Alaamian M et al (2004) The relationship of diaphragmatic defect, liver growth, and lung hypoplasia in nitrofen-induced congenital diaphragmatic hernia in the rat. Pediatr Surg Int 20:509–514

    Article  PubMed  Google Scholar 

  10. Tobin KA, Harsem NK, Dalen KT et al (2006) Regulation of ADRP expression by long-chain polyunsaturated fatty acids in BeWo cells, a human placental choriocarcinoma cell line. J Lipid Res 47:815–823

    Article  CAS  PubMed  Google Scholar 

  11. Friedmacher F, Fujiwara N, Hofmann AD et al (2014) Evidence for decreased lipofibroblast expression in hypoplastic rat lungs with congenital diaphragmatic hernia. Pediatr Surg Int 30:1023–1029

    Article  PubMed  Google Scholar 

  12. Shi HB, Yu K, Luo J et al (2015) Adipocyte differentiation-related protein promotes lipid accumulation in goat mammary epithelial cells. J Dairy Sci 98:6954–6964

    Article  CAS  PubMed  Google Scholar 

  13. Montedonico S, Nakazawa N, Puri P (2008) Congenital diaphragmatic hernia and retinoids: searching for an etiology. Pediatr Surg Int 24:755–761

    Article  PubMed Central  PubMed  Google Scholar 

  14. Noble BR, Babiuk RP, Clugston RD et al (2007) Mechanisms of action of the congenital diaphragmatic hernia-inducing teratogen nitrofen. Am J Physiol Lung Cell Mol Physiol 293:L1079–L1087

    Article  CAS  PubMed  Google Scholar 

  15. Kutasy B, Gosemann JH, Doi T et al (2012) Nitrofen interferes with trophoblastic expression of retinol-binding protein and transthyretin during lung morphogenesis in the nitrofen-induced congenital diaphragmatic hernia model. Pediatr Surg Int 28:143–148

    Article  PubMed  Google Scholar 

  16. Gosemann JH, Doi T, Kutasy B et al (2012) Alterations of peroxisome proliferator-activated receptor gamma and monocyte chemoattractant protein 1 gene expression in the nitrofen-induced hypoplastic lung. J Pediatr Surg 47:847–851

    Article  PubMed  Google Scholar 

  17. Kling DE, Cavicchio AJ, Sollinger CA et al (2010) Nitrofen induces apoptosis independently of retinaldehyde dehydrogenase (RALDH) inhibition. Birth Defects Res B Dev Reprod Toxicol 89:223–232

    CAS  PubMed  Google Scholar 

  18. Manson JM (1986) Mechanism of nitrofen teratogenesis. Environ Health Perspect 70:137–147

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  19. Brown TJ, Manson JM (1986) Further characterization of the distribution and metabolism of nitrofen in the pregnant rat. Teratology 34:129–139

    Article  CAS  PubMed  Google Scholar 

  20. Clugston RD, Zhang W, Greer JJ (2012) Early development of the primordial mammalian diaphragm and cellular mechanisms of nitrofen-induced congenital diaphragmatic hernia. Birth Defects Res A Clin Mol Teratol 88:15–24

    Google Scholar 

  21. Columbano A, Shinozuka H (1996) Liver regeneration versus direct hyperplasia. FASEB J 10:1118–1128

    CAS  PubMed  Google Scholar 

  22. Michalopoulos GK, DeFrances MC (1997) Liver regeneration. Science 276:60–66

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Our Lady’s Children’s Hospital, National Children’s Research Centre, Dublin 12, Ireland

    Hiromizu Takahashi, Balazs Kutasy, Florian Friedmacher, Toshiaki Takahashi & Prem Puri

Authors
  1. Hiromizu Takahashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Balazs Kutasy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Florian Friedmacher
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Toshiaki Takahashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Prem Puri
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Prem Puri.

Ethics declarations

Funding

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

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Takahashi, H., Kutasy, B., Friedmacher, F. et al. Expression of hepatic lipid droplets is decreased in the nitrofen model of congenital diaphragmatic hernia. Pediatr Surg Int 32, 155–160 (2016). https://doi.org/10.1007/s00383-015-3827-4

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00383-015-3827-4

Keywords

Search

Navigation