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Epimorphin expression in a rat model of pulmonary hypoplasia associated with congenital diaphragmatic hernia

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Abstract

Purpose

The pathogenesis of pulmonary hypoplasia associated with congenital diaphragmatic hernia (CDH) remains unclear. Interactions between the epithelium and surrounding mesenchyme play an important role in normal lung morphogenesis. Epimorphin, a stromal protein, plays a role in epithelial morphogenesis and lung branching, both of which are involved in pulmonary hypoplasia. In this study, we aimed to examine the relationship between epimorphin and pulmonary hypoplasia associated with CDH in an animal model.

Methods

Time-pregnant rats were exposed to nitrofen or vehicle on gestational day 9 (D9). Fetuses were harvested on D16 and D20, and were divided into control, hypoplastic lungs with CDH (CDH+), and hypoplastic lungs without CDH (CDH−). Both lungs of each fetus were removed and subjected to morphometric and molecular biologic analyses. Lung-to-body weight ratios were calculated. Pulmonary RNA was extracted, and relative mRNA level of epimorphin was determined by quantitative real-time PCR (qRT-PCR). Protein expression of epimorphin was investigated by Western blotting.

Results

In groups D16 and D20, lung-to-body weight ratios in subgroups CDH+ were significantly lower than those of controls and CDH−. The relative mRNA expression levels of epimorphin were significantly increased in both lungs in subgroup CDH+ compared with controls and CDH− on D16. Pulmonary epimorphin gene expression levels were significantly decreased in CDH+ group on D20 compared to controls. Western blotting confirmed the qRT-PCR results showing decreased pulmonary epimorphin protein expression in CDH+ hypoplastic lungs compared to controls on D20.

Conclusion

Our study shows that there is an association between the epimorphin expression and pulmonary hypoplasia associated with CDH. Although the cause–effect relationship is far from being established, epimorphin-related mechanisms have a more critical role in early (D16) developmental stage.

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Acknowledgments

The authors wish to thank Philip Jacobs for his suggestions on preparing the manuscript. This paper comprises the results of a project supported by The Scientific Research Projects Coordination Unit of Akdeniz University (Project No: 04.0103.005).

Conflict of interest

The authors report no conflict of interest.

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Authors and Affiliations

  1. Department of Pediatric Surgery, Akdeniz University Faculty of Medicine, Antalya, Turkey

    İlke Akpinar, Gungor Karaguzel, Cem Boneval & Mustafa Melikoglu

  2. Department of Biochemistry, Akdeniz University Faculty of Medicine, Antalya, Turkey

    Dicle Korgun, Arzu Çetin & Akin Yesilkaya

  3. Department of Gastroenterology, Institute of Health Economics, University of Alberta Faculty of Medicine, 1200 10405 Jasper Avenue, Edmonton, AB, T5J 3N4, Canada

    İlke Akpinar

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  1. İlke Akpinar
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Correspondence to İlke Akpinar.

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Akpinar, İ., Korgun, D., Çetin, A. et al. Epimorphin expression in a rat model of pulmonary hypoplasia associated with congenital diaphragmatic hernia. Pediatr Surg Int 30, 1037–1043 (2014). https://doi.org/10.1007/s00383-014-3579-6

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  • DOI: https://doi.org/10.1007/s00383-014-3579-6

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