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Nitro-Oleic Acid Prevents Hypoxia- and Asymmetric Dimethylarginine-Induced Pulmonary Endothelial Dysfunction

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Abstract

Rationale

Pulmonary hypertension (PH) represents a serious health complication accompanied with hypoxic conditions, elevated levels of asymmetric dimethylarginine (ADMA), and overall dysfunction of pulmonary vascular endothelium. Since the prevention strategies for treatment of PH remain largely unknown, our study aimed to explore the effect of nitro-oleic acid (OA-NO2), an exemplary nitro-fatty acid (NO2-FA), in human pulmonary artery endothelial cells (HPAEC) under the influence of hypoxia or ADMA.

Methods

HPAEC were treated with OA-NO2 in the absence or presence of hypoxia and ADMA. The production of nitric oxide (NO) and interleukin-6 (IL-6) was monitored using the Griess method and ELISA, respectively. The expression or activation of different proteins (signal transducer and activator of transcription 3, STAT3; hypoxia inducible factor 1α, HIF-1α; endothelial nitric oxide synthase, eNOS; intercellular adhesion molecule-1, ICAM-1) was assessed by the Western blot technique.

Results

We discovered that OA-NO2 prevents development of endothelial dysfunction induced by either hypoxia or ADMA. OA-NO2 preserves normal cellular functions in HPAEC by increasing NO production and eNOS expression. Additionally, OA-NO2 inhibits IL-6 production as well as ICAM-1 expression, elevated by hypoxia and ADMA. Importantly, the effect of OA-NO2 is accompanied by prevention of STAT3 activation and HIF-1α stabilization.

Conclusion

In summary, OA-NO2 eliminates the manifestation of hypoxia- and ADMA-mediated endothelial dysfunction in HPAEC via the STAT3/HIF-1α cascade. Importantly, our study is bringing a new perspective on molecular mechanisms of NO2-FAs action in pulmonary endothelial dysfunction, which represents a causal link in progression of PH.

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Acknowledgments

We would like to thank Jirina Prochazkova, Lenka Svihalkova Sindlerova, and Ondrej Vasicek for a fruitful discussion and insightful suggestions.

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Authors and Affiliations

  1. Institute of Biophysics, Academy of Sciences of the Czech Republic, v.v.i., Kralovopolska 135, 612 65, Brno, Czech Republic

    Adolf Koudelka, Gabriela Ambrozova, Tana Fidlerova, Hana Martiskova, Lukas Kubala & Michaela Pekarova

  2. International Clinical Research Center, St. Anne’s University Hospital, Pekarska 53, 656 91, Brno, Czech Republic

    Gabriela Ambrozova, Anna Klinke, Hana Martiskova, Lukas Kubala & Michaela Pekarova

  3. Department of Experimental Cardiology, University Hospital of Cologne, Kerpener Str. 62, 50924, Cologne, Germany

    Anna Klinke & Tanja K. Rudolph

  4. Faculty of Science, Institute of Experimental Biology, Masaryk University, Kamenice 5, 625 00, Brno, Czech Republic

    Tana Fidlerova & Hana Martiskova

  5. Faculty of Electrical Engineering and Communication, Brno University of Technology, Technicka 3058/10, 616 00, Brno, Czech Republic

    Radek Kuchta, Jaroslav Kadlec & Zdenka Kuchtova

  6. Department of Pharmacology and Chemical Biology, University of Pittsburgh, 4200 Fifth Ave, Pittsburgh, PA, 15260, USA

    Steven R. Woodcock & Bruce A. Freeman

Authors
  1. Adolf Koudelka
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  2. Gabriela Ambrozova
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  3. Anna Klinke
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  4. Tana Fidlerova
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  7. Tanja K. Rudolph
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  10. Steven R. Woodcock
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  11. Bruce A. Freeman
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  12. Lukas Kubala
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  13. Michaela Pekarova
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Corresponding author

Correspondence to Michaela Pekarova.

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Funding

This work was supported by the Czech Science Foundation (no. 13-40824P), the Ministry of Education, Youth and Sports (no. LD15069). LK and MP were supported by the European Regional Development Fund, the projects NPS II no. LQ1605 and FNUSA-ICRC no. CZ.1.05/1.1.00/02.0123 from MEYS CR. BAF was supported by NIH grants R01-HL-058115, R01-HL-64937, and PO1-HL-103455.

Conflict of Interest

Bruce A. Freeman (BAF) and Steven R. Woodcock (SRW) acknowledge an interest in Complexa, Inc., as scientific founder/shareholder (BAF) and consultant (SRW). All other authors declare no conflicts of interest with respect to the contents of this manuscript.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

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Koudelka, A., Ambrozova, G., Klinke, A. et al. Nitro-Oleic Acid Prevents Hypoxia- and Asymmetric Dimethylarginine-Induced Pulmonary Endothelial Dysfunction. Cardiovasc Drugs Ther 30, 579–586 (2016). https://doi.org/10.1007/s10557-016-6700-3

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