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Adenosine A2A receptor regulates expression of vascular endothelial growth factor in feto-placental endothelium from normal and late-onset pre-eclamptic pregnancies

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Abstract

We aim to investigate whether A2A/nitric oxide-mediated regulation of vascular endothelial growth factor (VEGF) expression is impaired in feto-placental endothelial cells from late-onset pre-eclampsia. Cultures of human umbilical vein endothelial cells (HUVECs) and human placental microvascular endothelial cells (hPMECs) from normal and pre-eclamptic pregnancies were used. Assays by using small interference RNA (siRNA) for A2A were performed, and transfected cells were used for estimation of messenger RNA (mRNA) levels of VEGF, as well as for cell proliferation and angiogenesis in vitro. CGS-21680 (A2A agonist, 24 h) increases HUVEC and hPMEC proliferation in a dose response manner. Furthermore, similar to CGS-21680, the nitric oxide donor, S-nitroso-N-acetyl-penicillamine oxide (SNAP), increased cell proliferation in a dose response manner (logEC50 10−9.2 M). In hPMEC, CGS-21680 increased VEGF protein levels in both normal (∼1.5-fold) and pre-eclamptic pregnancies (∼1.2-fold), an effect blocked by the A2A antagonist, ZM-241385 (10−5 M) and the inhibitor of NO synthase, N ω-nitro-L-arginine methyl ester hydrochloride (L-NAME). Subsequently, SNAP partially recovered cell proliferation and in vitro angiogenesis capacity of cells from normal pregnancies exposed to siRNA for A2A. CGS-21680 also increased (∼1.5-fold) the level of VEGF mRNA in HUVEC from normal pregnancies, but not in pre-eclampsia. Additionally, transfection with siRNA for A2A decrease (∼30 %) the level of mRNA for VEGF in normal pregnancy compared to untransfected cells, an effect partially reversed by co-incubation with SNAP. The A2A-NO-VEGF pathway is present in endothelium from microcirculation and macrocirculation in both normal and pre-eclamptic pregnancies. However, NO signaling pathway seems to be impaired in HUVEC from pre-eclampsia.

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Abbreviations

CGS-21680:

2-(p-(2-Carbonyl-ethyl)-feniletilamino)-50-N-ethylcarboxamidoadenosine)

BrdU:

5-Bromo-2-deoxyuridine

SCH-58261:

7-(2-Phenylethyl)-5-amino-2-(2-furyl)-pyrazolo-(4,3-e)-1,2,4-triazolo(1,5-c)pyrimidine

AR:

Adenosine receptors

HMEC-1:

Dermal microvascular endothelial cells

EOPE:

Early onset pre-eclampsia

hPMEC:

Human placental microvascular endothelial cells

HUVEC:

Human umbilical vein endothelial cells

LOPE:

Late-onset pre-eclampsia

NECA:

N-ethylcarboxamidoadenosine

NO:

Nitric oxide

L-NAME:

N ω-nitro-L-arginine methyl ester hydrochloride

VEGF:

Vascular endothelial growth factor

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Acknowledgments

We would like to thank all research staff at the Vascular Physiology Laboratory and the Group of Investigation of Tumor Angiogenesis (GIANT) of the Universidad del Bío-Bío for their technical support. We also give thanks to researchers in the GRIVAS Health group for outstanding discussion regarding ideas presented in this manuscript. We thank Dr. Igor Feoktistov from University of Vanderbilt for active discussion and molecular tools kindly shared with our lab. This study was financed by Fondecyt Regular 1140586, Fondequip EQM140104, DIUBB GI153109/EF, and GI 152920/EF.

Individual contributions

This work was carried out as a full collaboration among all the authors. CE defined the research topic. JA, KH, FT, and CA prepared draft of the manuscript. PB is the clinical responsible for patient inclusion. CE, JA, and CA co-wrote the manuscript. All authors approved the final version of the manuscript.

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Authors and Affiliations

  1. Vascular Physiology Laboratory, Group of Investigation in Tumor Angiogenesis (GIANT), Department of Basic Sciences, Universidad del Bío-Bío, Chillán, Chile

    Jesenia Acurio, Kurt Herlitz, Felipe Troncoso & Carlos Escudero

  2. Department of Clinical Biochemistry and Immunology, Faculty of Pharmacy, University of Concepción, Concepción, Chile

    Claudio Aguayo

  3. Group of Research and Innovation in Vascular Health (GRIVAS Health), Chillan, Chile

    Claudio Aguayo & Carlos Escudero

  4. Obstetric and Gynecology Department, Hospital Clinico Herminda Martin, Chillán, Chile, Universidad Católica de la Santísima Concepción, Concepción, Chile

    Patricio Bertoglia

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  1. Jesenia Acurio
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  2. Kurt Herlitz
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  3. Felipe Troncoso
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  4. Claudio Aguayo
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  6. Carlos Escudero
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Correspondence to Carlos Escudero.

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Acurio, J., Herlitz, K., Troncoso, F. et al. Adenosine A2A receptor regulates expression of vascular endothelial growth factor in feto-placental endothelium from normal and late-onset pre-eclamptic pregnancies. Purinergic Signalling 13, 51–60 (2017). https://doi.org/10.1007/s11302-016-9538-z

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