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. Graphical Abstract ᅟ.

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

Department of Pharmacology and Chemical Biology University of Pittsburgh 4200 5th Ave Pittsburgh PA 15260 USA

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

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

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

International Clinical Research Center St Anne's University Hospital Pekarska 53 656 91 Brno Czech Republic

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Sitbon O, Lascoux-Combe C, Delfraissy JF, et al. Prevalence of HIV-related pulmonary arterial hypertension in the current antiretroviral therapy era. Am J Resp Crit Care. 2008;177:108–13. PubMed

Schermuly RT, Ghofrani HA, Wilkins MR, Grimminger F. Mechanisms of disease: pulmonary arterial hypertension. Nat Rev Cardiol. 2011;8:443–55. PubMed PMC

Fonseca GH, Souza R, Salemi VM, Jardim CV, Gualandro SF. Pulmonary hypertension diagnosed by right heart catheterization in sickle cell disease. Eur Respir J. 2012;39:112–8. PubMed

Sanli C, Oguz D, Olgunturk R, et al. Elevated homocysteine and asymmetric dimethyl arginine levels in pulmonary hypertension associated with congenital heart disease. Pediatr Cardiol. 2012;33:1323–31. PubMed

Lüneburg N, Harbaum L, Hennigs JK. The endothelial ADMA/NO pathway in hypoxia-related chronic respiratory diseases. Biomed Res Int. 2014 PubMed PMC

Parikh RV, Scherzer R, Nitta EM, et al. Increased levels of asymmetric dimethylarginine are associated with pulmonary arterial hypertension in HIV infection. AIDS. 2014;28:511–9. PubMed PMC

Trittmann JK, Peterson E, Rogers LK, et al. Plasma asymmetric dimethylarginine levels are increased in neonates with bronchopulmonary dysplasia-associated pulmonary hypertension. J Pediatr. 2015;166:230–3. PubMed PMC

Iannone L, Zhao L, Dubois O, et al. miR-21/DDAH1 pathway regulates pulmonary vascular responses to hypoxia. Biochem J. 2014;462:103–12. PubMed

Pekarova M, Koudelka A, Kolarova H, et al. Asymmetric dimethyl arginine induces pulmonary vascular dysfunction via activation of signal transducer and activator of transcription 3 and stabilization of hypoxia-inducible factor 1-alpha. Vasc Pharmacol. 2015;73:138–48. PubMed

Rudolph TK, Rudolph V, Edreira MM, et al. Nitro-fatty acids reduce atherosclerosis in apolipoprotein E-deficient mice. Arterioscl Throm Vas. 2010;30:938–45. PubMed PMC

Rudolph V, Rudolph TK, Schopfer FJ, et al. Endogenous generation and protective effects of nitro-fatty acids in a murine model of focal cardiac ischaemia and reperfusion. Cardiovasc Res. 2010;85:155–66. PubMed PMC

Khoo NK, Rudolph V, Cole MP, et al. Activation of vascular endothelial nitric oxide synthase and heme oxygenase-1 expression by electrophilic nitro-fatty acids. Free Radic Biol Med. 2010;48:230–9. PubMed PMC

Zhang J, Villacorta L, Chang L, et al. Nitro-oleic acid inhibits angiotensin II–induced hypertension. Circ Res. 2010;107:540–8. PubMed PMC

Kansanen E, Jyrkkänen HK, Volger OL, et al. Nrf2-dependent and -independent responses to nitro-fatty acids in human endothelial cells: identification of heat shock response as the major pathway activated by nitro-oleic acid. J Biol Chem. 2009;284:33233–41. PubMed PMC

Schopfer FJ, Cole MP, Groeger AL, et al. Covalent peroxisome proliferator-activated receptor{gamma} binding by nitro-fatty acids: endogenous ligands act as selective modulators. J Biol Chem. 2010;285:12321–33. PubMed PMC

Klinke A, Möller A, Pekarova M, et al. Protective effects of 10-nitro-oleic acid in a hypoxia-induced murine model of pulmonary hypertension. Am J Resp Cell Mol. 2014;51:155–62. PubMed PMC

Yang XP, Irani K, Mattagajasingh S, et al. Signal transducer and activator of transcription 3alpha and specificity protein 1 interact to upregulate intercellular adhesion molecule-1 in ischemic-reperfused myocardium and vascular endothelium. Arterioscl Throm Vasc. 2005;25:1395–400. PubMed

Paulin R, Meloche J, Bonnet S. STAT3 signaling in pulmonary arterial hypertension. JAK-STAT. 2012;1:223–33. PubMed PMC

Sun CK, Zhen YY, Lu HI, Sung PH, et al. Reducing TRPC1 expression through liposome-mediated siRNA delivery markedly attenuates hypoxia-induced pulmonary arterial hypertension in a murine model. Stem Cells Int. 2014;2014:316214. PubMed PMC

Ambrozova G, Martiskova H, Koudelka A, et al. Nitro-oleic acid modulates classical and regulatory activation of macrophages and their involvement in pro-fibrotic responses. Free radical bio med. 2016;90:252–60. PubMed PMC

Lim DG, Sweeney S, Bloodsworth a, et al. Nitrolinoleate, a nitric oxide-derived mediator of cell function: synthesis, characterization, and vasomotor activity. P Natl Acad Sci USA. 2002;99:15941–6. PubMed PMC

Lima ES, Bonini MG, Augusto O, Barbeiro HV, Souza HP, Abdalla DS. Nitrated lipids decompose to nitric oxide and lipid radicals and cause vasorelaxation. Free Radic Biol Med. 2005;39:532–9. PubMed

Freeman BA, Baker PR, Schopfer FJ, Woodcock SR, Napolitano A, d’Ischia M. Nitro-fatty acid formation and signaling. J Biol Chem. 2008;283:15515–9. PubMed PMC

Shin E, Yeo E, Lim J, et al. Nitrooleate mediates nitric oxide synthase activation in endothelial cells. Lipids. 2014;49:457–66. PubMed

Wang H, Jia Z, Sun J, et al. Nitrooleic acid protects against cisplatin nephropathy: role of COX-2/mPGES-1/PGE2 cascade. Mediat Inflamm. 2015 PubMed PMC

Balazy M, Iesaki T, Park JL, et al. Vicinal nitrohydroxyeicosatrienoic acids: vasodilator lipids formed by reaction of nitrogen dioxide with arachidonic acid. J Pharmacol Exp Ther. 2001;299:611–9. PubMed

Schopfer FJ, Baker PR, Giles G, et al. Fatty acid transduction of nitric oxide signaling. Nitrolinoleic acid is a hydrophobically stabilized nitric oxide donor. J Biol Chem. 2005;280:19289–97. PubMed

Ambrozova G, Fidlerova T, Verescakova H, et al. Nitro-oleic acid inhibits vascular endothelial inflammatory responses and the endothelial-mesenchymal transition. Biochim Biophys Acta. 2016 pii: S0304–4165(16)30251–30253. PubMed PMC

Reddy AT, Lakshmi SP, Dornadula S, Pinni S, Rampa DR, Reddy RC. The nitrated fatty acid 10-nitro-oleate attenuates allergic airway disease. J Immunol. 2013;191:2053–63. PubMed

Villacorta L, Chang L, Salvatore SR, et al. Electrophilic nitro-fatty acids inhibit vascular inflammation by disrupting LPS-dependent TLR4 signalling in lipid rafts. Cardiovasc Res. 2013;98:116–24. PubMed PMC

Wu Y, Dong Y, Song P, Zou MH. Activation of the AMP-activated protein kinase (AMPK) by nitrated lipids in endothelial cells. PLoS One. 2012 PubMed PMC

Stenmark KR, Fagan KA, Frid MG. Hypoxia-induced pulmonary vascular remodeling: cellular and molecular mechanisms. Circ Res. 2006;99:675–91. PubMed

Rudnicki M, Faine LA, Dehne N, et al. Hypoxia inducible factor-dependent regulation of angiogenesis by nitro-fatty acids. Arterioscl Throm Vasc. 2011;31:1360–7. PubMed

Saura M, Zaragoza C, Bao C, Herranz B, Rodriguez-Puyol M, Lowenstein CJ. Stat3 mediates interleukin-6 [correction of interelukin-6] inhibition of human endothelial nitric-oxide synthase expression. J Biol Chem. 2006;281:30057–62. PubMed