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Congenital diaphragmatic hernia: phosphodiesterase-5 and Arginase inhibitors prevent pulmonary vascular hypoplasia in rat lungs

Pediatric Research volume 95, pages 941–948 (2024)Cite this article

Abstract

Background

Severe pulmonary hypoplasia related to congenital diaphragmatic hernia (CDH) continues to be a potentially fatal condition despite advanced postnatal management strategies.

Objective

To evaluate the effect of the antenatal sildenafil and 2(S)-amino-6-boronohexanoic acid (ABH-Arginase inhibitor) on lung volume, pulmonary vascular development, and nitric oxide (NO) synthesis in a Nitrofen-induced CDH rat model.

Methods

Nitrofen-induced CDH rat model was used. Nitrofen was administrated on embryonic day(E) 9,5. At E14, five intervention groups were treated separately: Nitrofen, Nitrofen+Sildenafil, Nitrofen+ABH, Nitrofen+Sildenafil+ABH and Control. At term, offspring’s lungs were weighed, some paraffin-embedded for histology, others snap-frozen to analyze eNOS, Arginase I–II expression, and activity.

Results

In CDH-bearing offsprings, ABH or Sildenafil+ABH preserved the total lung/body-weight index (p < 0.001), preventing pulmonary vascular smooth muscle cell hyperproliferation and improving lung morphometry. Sildenafil+ABH increased 1.7-fold the lung nitrite levels (p < 0.01) without changes in eNOS expression. Sildenafil and ABH improved the number of pulmonary vessels.

Conclusion

These results suggest that in this CDH rat model, the basal activity of Arginase participates in the lung volume and, together with phosphodiesterase-5, regulates NOS activity in the term fetal lung. The combined treatment (Sildenafil+ABH) could revert some of the pulmonary features in CDH by improving the local NO synthesis and preventing smooth muscle cell hyperproliferation.

Impact

  • This study presents Arginase inhibition as a new therapeutic target and the importance of the combined antenatal treatment to improve pulmonary vascular development in a congenital diaphragmatic hernia (CDH) rat model.

  • This study shows that the action of an Arginase inhibitor (ABH) enhances the effects already described for sildenafil in this model.

  • These results reinforce the importance of prenatal treatments’ synergy in recovering the hypoplastic lung in the Nitrofen-induced CDH rat model.

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Fig. 1: Protocol overview.
Fig. 2: Morphometry of lung size, vascularity and muscular wall thickness.
Fig. 3: Histological characteristics of pulmonary vessels and tissue in controls and different treatment groups.
Fig. 4: Nitrites and eNOS and Arginase expression in lung tissue of the offspring.
Fig. 5: Effect of the combination of prenatal treatment with ABH and Sildenafil in the CDH nitrofen-induced rat model.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This research was supported by the following grants: FONDECYT N° 1171406 (P.C.) and Grant PIA CONICYT ACE210009 to CARE-UC and a donation of SQM to the Pontificia Universidad Católica de Chile (PUC). The UC CINBIOT Animal Facility is funded by PIA CONICYT ECM-07. Grant Faculty of Medicine, Pediatric Division at PUC. We thank MECESUP PUC0815 grant – Equipamiento Científico Mayor Centro de Investigaciones Médicas of Pontificia Universidad Católica de Chile for the access to Synergy II microplate fluorescence reader (BioTek Instruments, Winooski, VT) and Carl Zeiss Axio Imager A1 fluorescence microscope (Göttingen, Germany).

Funding

Fondo Nacional de Desarrollo Científico y Tecnológico (FONDECYT) #1171406, Grant PIA CONICYT AFB170005, Proyecto Semilla Interdisciplinario PS 14/15, DIDEMUC, Faculty of Medicine and Research grant from the Pediatric Division at PUC.

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Author notes

  1. Carlos Céspedes & Carlos P. Vio

    Present address: Faculty of Medicine and Science, Universidad San Sebastián, Santiago, Chile

Authors and Affiliations

  1. Department of Neonatology, Pontificia Universidad Católica de Chile, Santiago, Chile

    Alberto Toso, Oscar Aránguiz, Cherie Hernández, Matías Luco, Paola Casanello & Javier Kattan

  2. Department of Obstetrics, Pontificia Universidad Católica de Chile, Santiago, Chile

    Oscar Aránguiz, Cherie Hernández & Paola Casanello

  3. Center for Aging and Regeneration CARE UC, Department of Physiology, Faculty of Biological Sciences, Pontificia Universidad Católica de Chile, Santiago, Chile

    Carlos Céspedes & Carlos P. Vio

  4. Department of Pathology, Pontificia Universidad Católica de Chile, Santiago, Chile

    Orieta Navarrete

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Contributions

J.K., P.C., and A.T. conceived and designed the experiments. A.T., O.A., C.C., C.P.V., and C.H. collected and analyzed the experimental data. J.K., P.C., A.T., O.A., and O.N. interpreted the experimental data. J.K., P.C., A.T., and O.A. drafted the article. All authors critically read and corrected the manuscript draft and approved the final manuscript.

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Correspondence to Paola Casanello or Javier Kattan.

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Toso, A., Aránguiz, O., Céspedes, C. et al. Congenital diaphragmatic hernia: phosphodiesterase-5 and Arginase inhibitors prevent pulmonary vascular hypoplasia in rat lungs. Pediatr Res 95, 941–948 (2024). https://doi.org/10.1038/s41390-022-02366-4

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