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miR-1 induces endothelial dysfunction in rat pulmonary arteries

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Abstract

Endothelial dysfunction plays a central role in the pathophysiology of pulmonary arterial hypertension (PAH). MicroRNAs (miRNAs) are small single-strand and non-coding RNAs that negatively regulate gene function by binding to the 3′-untranslated region (3′-UTR) of specific mRNAs. microRNA-1 (miR-1) is upregulated in plasma from idiopathic PAH patients and in lungs from an experimental model of PAH. However, the role of miRNA-1 on endothelial dysfunction is unknown. The aim of this study was to analyze the effects of miR-1 on endothelial function in rat pulmonary arteries (PA). Endothelial function was studied in PA from PAH or healthy animals and mounted in a wire myograph. Some PA from control animals were transfected with miR-1 or scramble miR. Superoxide anion production by miR-1 was quantified by dihydroethidium (DHE) fluorescence in rat PA smooth muscle cells (PASMC). Bioinformatic analysis identified superoxide dismutase-1 (SOD1), connexin-43 (Cx43), caveolin 2 (CAV2) and Krüppel-like factor 4 (KLF4) as potential targets of miR-1. The expression of SOD1, Cx43, CAV2, and KLF4 was determined by qRT-PCR and western blot in PASMC. PA incubated with miR-1 presented decreased endothelium-dependent relaxation to acetylcholine. We also found an increase in the production of O2− and decreased expression of SOD1, Cx43, CAV2, and KLF4 in PASMC induced by miR-1, which may contribute to endothelial dysfunction. In conclusion, these data indicate that miR-1 induces endothelial dysfunction, suggesting a pathophysiological role in PAH.

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Funding

Authors research is funded by Ministerio de Economía y Competitividad grants (SAF2014–55399-R, SAF2016-77222R), Comunidad de Madrid (B2017/BMD-3727), Instituto de Salud Carlos III (PI15/01100), and Fundación Contra la Hipertensión Pulmonar (Empathy) with funds from the European Union (Fondo Europeo de Desarrollo Regional FEDER). G.M-P, M.C., and S.E-R. are funded by Ciberes grant with funds from Fundación Contra la Hipertensión Pulmonar, UCM predoctoral grant, and a FPU grant from Ministerio de Educación, respectively.

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Author notes

  1. Gema Mondejar-Parreño, María Callejo, Angel Cogolludo and Francisco Perez-Vizcaino contributed equally to this work.

Authors and Affiliations

  1. Departament of Pharmacology and Toxicology. School of Medicine, Universidad Complutense de Madrid, 28040, Madrid, Spain

    Gema Mondejar-Parreño, María Callejo, Bianca Barreira, Daniel Morales-Cano, Sergio Esquivel-Ruiz, Laura Moreno, Angel Cogolludo & Francisco Perez-Vizcaino

  2. Ciber Enfermedades Respiratorias (Ciberes), Madrid, Spain

    Gema Mondejar-Parreño, María Callejo, Bianca Barreira, Daniel Morales-Cano, Sergio Esquivel-Ruiz, Marco Filice, Laura Moreno, Angel Cogolludo & Francisco Perez-Vizcaino

  3. Instituto de Investigación Sanitaria Gregorio Marañón (IISGM), Madrid, Spain

    Gema Mondejar-Parreño, María Callejo, Bianca Barreira, Daniel Morales-Cano, Sergio Esquivel-Ruiz, Laura Moreno, Angel Cogolludo & Francisco Perez-Vizcaino

  4. Department of Chemistry in Pharmaceutical Sciences, Faculty of Pharmacy, Universidad Complutense de Madrid, Madrid, Spain

    Marco Filice

  5. Departamento de Farmacología y Toxicología, Facultad de Medicina, Universidad Complutense de Madrid, 28040, Madrid, Spain

    Francisco Perez-Vizcaino

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  1. Gema Mondejar-Parreño
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Contributions

GMP, MC, BB, DMC, and SE performed and analyzed the experiments. GMP and MC drafted the manuscript. AC and FPV conceived the study and designed the experiments. FPV wrote the manuscript with significant conceptual contributions from GMP, MC, LM, MF, and AC.

Corresponding author

Correspondence to Francisco Perez-Vizcaino.

Ethics declarations

All experimental procedures utilizing animals were carried out according to the Care and Use of Laboratory Animals and approved by the institutional Ethical Committees of the Universidad Complutense de Madrid (Madrid, Spain) and the regional Committee for Laboratory Animals Welfare (Comunidad de Madrid, Ref. number PROEX-251/15). All investigators understand the ethical principles.

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Mondejar-Parreño, G., Callejo, M., Barreira, B. et al. miR-1 induces endothelial dysfunction in rat pulmonary arteries. J Physiol Biochem 75, 519–529 (2019). https://doi.org/10.1007/s13105-019-00696-2

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