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The P2Y1 receptor-mediated leukocyte adhesion to endothelial cells is inhibited by melatonin

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Abstract

Extracellular ATP (released by endothelial and immune cells) and its metabolite ADP are important pro-inflammatory mediators via the activation of purinergic P2 receptors (P2Y and P2X), which represent potential new targets for anti-inflammatory therapy. Endothelial P2Y1 receptor (P2Y1R) induces endothelial cell activation triggering leukocyte adhesion. A number of data have implicated melatonin as a modulator of immunity, inflammation, and endothelial cell function, but to date no studies have investigated whether melatonin modulates endothelial P2YR signaling. Here, we evaluated the putative effect of melatonin on P2Y1R-mediated leukocyte adhesion to endothelial cells and TNF-α production, using mesenteric endothelial cells and fresh peripheral blood mononuclear cells isolated from rats. Endothelial cells were treated with the P2Y1R agonist 2MeSATP, alone or in combination with melatonin, and then exposed to mononuclear cells. 2MeSATP increased leukocyte adhesion to endothelial cells and TNF-α production in vitro, and melatonin inhibited both effects without altering P2Y1R protein expression. In addition, assays with the Ca2+ chelator BAPTA-AM indicate that the effect of melatonin on 2MeSATP-stimulated leukocyte adhesion depends on intracellular Ca2+ modulation. P2Y1R is considered a potential target to control chronic inflammation. Therefore, our data unveiled a new endothelial cell modulator of purinergic P2Y1 receptor signaling.

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Acknowledgements

CLMS is senior fellow of CNPq (Brazil). The authors thank Orlando da Rocha Moreira (UFRJ) for technical assistance.

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Authors and Affiliations

  1. Laboratory of Biochemical and Molecular Pharmacology, Institute of Biomedical Sciences, Federal University of Rio de Janeiro, Av. Carlos Chagas Filho, 373 CCS room J-17, Cidade Universitária CEP 21941-599, Rio de Janeiro, RJ, 21941-902, Brazil

    Tassya Cataldi Cardoso, Thaís Emanuelle Pompeu & Claudia Lucia Martins Silva

Authors
  1. Tassya Cataldi Cardoso
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  2. Thaís Emanuelle Pompeu
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  3. Claudia Lucia Martins Silva
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Correspondence to Claudia Lucia Martins Silva.

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Funding

This study was funded by National Council for Scientific and Technological Development (CNPq, Brazil, grant number 455436/2014-2).

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Tassya Cataldi Cardoso declares that she has no conflict of interest.

Thaís Emanuelle Pompeu declares that she has no conflict of interest.

Claudia Lucia Martins Silva declares that she has no conflict of interest.

Ethical approval

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

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Cardoso, T.C., Pompeu, T.E. & Silva, C.L.M. The P2Y1 receptor-mediated leukocyte adhesion to endothelial cells is inhibited by melatonin. Purinergic Signalling 13, 331–338 (2017). https://doi.org/10.1007/s11302-017-9565-4

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  • DOI: https://doi.org/10.1007/s11302-017-9565-4

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