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Downregulation of Midkine gene expression and its response to retinoic acid treatment in the nitrofen-induced hypoplastic lung

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Abstract

Purpose

Nitrofen-induced congenital diaphragmatic hernia (CDH) model has been widely used to investigate the pathogenesis of pulmonary hypoplasia (PH) in CDH. Recent studies have suggested that retinoids may be involved in the molecular mechanisms of PH in CDH. Prenatal treatment with retinoic acid (RA) has been reported to improve the growth of hypoplastic lung in the nitrofen CDH model. Midkine (MK), a RA-responsive growth factor, plays key roles in various organogenesis including lung development. In fetal lung, MK mRNA expression has its peak at E13.5–E16.5 and is markedly decreased during mid-to-late gestation, indicating its important role in early lung morphogenesis. We designed this study to investigate the hypothesis that the pulmonary MK gene expression is downregulated in the early lung morphogenesis in the nitrofen-induced PH, and to evaluate the effect of prenatal RA treatment on pulmonary MK gene expression in the nitrofen-induced CDH model.

Methods

Pregnant rats were exposed to either olive oil or nitrofen on day 9 of gestation (D9). Fetal lungs were harvested on D15, D18, and D21 and divided into control, nitrofen with or without CDH [CDH(+) or CDH(−)]. In addition, RA was given on days D18, D19, and D20 and fetal lungs were harvested on D21, and then divided into control + RA and nitrofen + RA. The pulmonary gene expression levels of MK were evaluated by real-time RT-PCR and statistically analyzed. Immunohistochemistry was also performed to examine protein expression/distribution of MK in fetal lung.

Results

The relative mRNA expression levels of MK were significantly downregulated in nitrofen group compared to controls at D15 (§ p < 0.01), whereas there were no significant differences at D18 and D21. MK gene expression levels were significantly upregulated in nitrofen + RA (0.71 ± 0.17) compared to the control (0.35 ± 0.16), CDH(−) (0.24 ± 0.15), CDH(+) (0.39 ± 0.19) and control + RA (0.47 ± 0.13) (*p < 0.05). Immunoreactivity of MK was also markedly decreased in nitrofen lungs compared to controls on D15, and increased in nitrofen + RA lungs compared to the other lungs on D21.

Conclusion

Downregulation of MK gene on D15 may contribute to primary PH in the nitrofen CDH model by disrupting early lung morphogenesis. Upregulation of MK gene after RA treatment in the nitrofen-induced hypoplastic lung suggests that RA may have a therapeutic potential to rescue PH in CDH through RA-responsive growth factor signaling.

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References

  1. Doyle NM, Lally KP (2004) The CDH study group and advance in the clinical care of the patient with congenital diaphragmatic hernia. Semin Perinatol 28:174–184

    Article  PubMed  Google Scholar 

  2. Grosche JR, Islam S, Boulanger SC (2005) Congenital diaphragmatic hernia. Am J Surg 190:324–332

    Article  Google Scholar 

  3. Kluth D, Kangah R, Reich P et al (1990) Nitrofen-induced diaphragmatic hernias in rats: an animal model. J Pediatr Surg 25:850–854

    Article  CAS  PubMed  Google Scholar 

  4. Ruttenstock E, Doi T, Dingemann J et al (2010) Downregulation of insulin-like growth factor binding protein 3 and 5 in nitrofen-induced pulmonary hypoplasia. Pediatr Surg Int 26:59–63

    Article  PubMed  Google Scholar 

  5. Doi T, Puri P (2009) Up-regulation of Wnt5a gene expression in the nitrofen-induced hypoplastic lung. J Pediatr Surg 44:2302–2306

    Article  PubMed  Google Scholar 

  6. Doi T, Hajduk P, Puri P (2009) Upregulation of Slit-2 and Slit-3 gene expressions in the nitrofen-induced hypoplastic lung. J Pediatr Surg 44:2092–2095

    Article  PubMed  Google Scholar 

  7. McCaffery PJ, Drager UC et al (2000) Retinoids in embryonal development. Physiol Rev 80:1021–1054

    PubMed  Google Scholar 

  8. Maden M (2004) Retinoids in lung development and regeneration. Curr Top Dev Biol 61:153–189

    Article  CAS  PubMed  Google Scholar 

  9. 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 293:1079–1087

    Article  Google Scholar 

  10. Nakazawa N, Montedonico S, Takayasu H et al (2007) Disturbance of retinol transportation causes nitrofen induced hypoplastic lung. J Pediatr Surg 42:345–349

    Article  PubMed  Google Scholar 

  11. Montedonico S, Nakazawa N, Puri P (2006) Retinoic acid rescues lung hypoplasia in nitrofen-induced hypoplastic foetal rat lung explants. Pediatr Surg Int 22:2–8

    Article  PubMed  Google Scholar 

  12. Montedonico S, Sugimoto K, Felle P et al (2008) Prenatal treatment with retinoic acid promotes pulmonary alveologenesis in the nitrofen model of congenital diaphragmatic hernia. J Pediatr Surg 43:500–507

    Article  PubMed  Google Scholar 

  13. Doi T, Sugimoto K, Puri P (2009) Prenatal retinoic acid up-regulates pulmonary gene expression of COUP-TFII, FOG2, and GATA4 in pulmonary hypoplasia. J Pediatr Surg 44:1933–1937

    Article  PubMed  Google Scholar 

  14. Doi T, Sugimoto K, Ruttenstock E et al (2010) Prenatal retinoic acid upregulates pulmonary gene expression of PI3K and AKT in nitrofen-induced pulmonary hypoplasia. Pediatr Surg Int 26(10):1011–1015

    Article  PubMed  Google Scholar 

  15. Zhang H, Garber SJ, Cui Z et al (2009) The angiogenic factor midkine is regulated by dexamethasone and retinoic acid during alveolarization and in alveolar epithelial cells. Respir Res 10:77

    Article  PubMed  Google Scholar 

  16. Kadomatsu K, Huang RP, Suganuma T et al (1990) A retinoic acid responsive gene MK found in the teratocarcinoma system is expressed in spatially and temporally controlled manner during mouse embryogenesis. J Cell Biol 110:607–616

    Article  CAS  PubMed  Google Scholar 

  17. Reynolds PR, Mucenski ML, Whitsett JA (2003) Thyroid transcription factor (TTF)-1 regulates the expression of midkine (MK) during lung morphogenesis. Dev Dyn 227:227–237

    Article  CAS  PubMed  Google Scholar 

  18. Kaplan F, Comber J, Sladek R et al (2003) The growth factor midkine is modulated by both glucocorticoid and retinoid in fetal lung development. Am J Respir Cell Mol Biol 28:33–41

    Article  CAS  PubMed  Google Scholar 

  19. Major D, Cadenas M, Fournier L et al (1998) Retinol status of newborn infants with congenital diaphragmatic hernia. Pediatr Surg Int 13:547–549

    Article  CAS  PubMed  Google Scholar 

  20. Massaro GD, Massaro D (1996) Postnatal treatment with retinoic acid increases the number of pulmonary alveoli in rats. Am J Physiol 270:L305–L310

    CAS  PubMed  Google Scholar 

  21. Hind M, Maden M (2004) Retinoic acid induces alveolar regeneration in the adult mouse lung. Eur Respir J 23:20–27

    Article  CAS  PubMed  Google Scholar 

  22. Mitsiadis TA, Muramatsu T, Muramatsu H et al (1995) Midkine (MK), a heparin-binding growth/differentiation factor, is regulated by retinoic acid and epithelial–mesenchymal interactions in the developing mouse tooth, and affects cell proliferation and morphogenesis. J Cell Biol 129:267–281

    Article  CAS  PubMed  Google Scholar 

  23. Oshika E, Liu S, Singh G et al (1998) Antagonistic effects of dexamethasone and retinoic acid on rat lung morphogenesis. Pediatr Res 43:315–324

    Article  CAS  PubMed  Google Scholar 

  24. Lazzaro D, Price M, de Felice M et al (1991) The transcription factor TTF-1 is expressed at the onset of thyroid and lung morphogenesis and in restricted regions of the foetal brain. Development 113:1093–1104

    CAS  PubMed  Google Scholar 

  25. Zhou L, Lim L, Costa RH et al (1996) Thyroid transcription factor-1, hepatocyte nuclear factor-3beta, surfactant protein B, C, and Clara cell secretory protein in developing mouse lung. J Histochem Cytochem 44:1183–1193

    CAS  PubMed  Google Scholar 

  26. Kimura S, Hara Y, Pineau T et al (1996) The T/ebp null mouse: thyroid-specific enhancer-binding protein is essential for the organogenesis of the thyroid, lung, ventral forebrain, and pituitary. Genes Dev 10:60–69

    Article  CAS  PubMed  Google Scholar 

  27. Losada A, Xia H, Migliazza L et al (1999) Lung hypoplasia caused by nitrofen is mediated by down-regulation of thyroid transcription factor TTF-1. Pediatr Surg Int 15:188–191

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

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

    Takashi Doi, Mika Shintaku, Jens Dingemann, Elke Ruttenstock & Prem Puri

  2. School of Medicine and Medical Science, Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Dublin, Ireland

    Takashi Doi & Prem Puri

Authors
  1. Takashi Doi
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  2. Mika Shintaku
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  3. Jens Dingemann
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  4. Elke Ruttenstock
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  5. Prem Puri
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Correspondence to Prem Puri.

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Doi, T., Shintaku, M., Dingemann, J. et al. Downregulation of Midkine gene expression and its response to retinoic acid treatment in the nitrofen-induced hypoplastic lung. Pediatr Surg Int 27, 199–204 (2011). https://doi.org/10.1007/s00383-010-2773-4

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  • DOI: https://doi.org/10.1007/s00383-010-2773-4

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