Prostacyclin prevents nitric oxide-induced megakaryocyte apoptosis

1. We have previously demonstrated that nitric oxide (NO) triggers CD34⁺-derived megakaryocyte apoptosis. We here show that prostacyclin (PGI₂) inhibits PAPA/NO-induced megakaryocyte death detected by fluorescent microscopy and flow cytometry. 2. The cAMP-specific phosphodiesterase inhibitor, Ro 20...

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Autores principales: Pozner, Roberto Gabriel, Negrotto, Soledad, D'Atri, Lina Paola, Kotler, Mónica Lidia, Lazzari, María Angela, Gómez, Ricardo Martín, Schattner, Mirta
Formato: Articulo
Lenguaje:Inglés
Publicado: 2005
Materias:
NO
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/127155
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id I19-R120-10915-127155
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Exactas
Biología
PGI₂
NO
cAMP
cGMP
caspase
spellingShingle Ciencias Exactas
Biología
PGI₂
NO
cAMP
cGMP
caspase
Pozner, Roberto Gabriel
Negrotto, Soledad
D'Atri, Lina Paola
Kotler, Mónica Lidia
Lazzari, María Angela
Gómez, Ricardo Martín
Schattner, Mirta
Prostacyclin prevents nitric oxide-induced megakaryocyte apoptosis
topic_facet Ciencias Exactas
Biología
PGI₂
NO
cAMP
cGMP
caspase
description 1. We have previously demonstrated that nitric oxide (NO) triggers CD34⁺-derived megakaryocyte apoptosis. We here show that prostacyclin (PGI₂) inhibits PAPA/NO-induced megakaryocyte death detected by fluorescent microscopy and flow cytometry. 2. The cAMP-specific phosphodiesterase inhibitor, Ro 20-1724, and the permeable analog dibutyryl-cAMP also delayed apoptosis. PGI₂ effect was fully prevented when adenylyl cyclase activity was suppressed by SQ 22536, and partially reversed by the permeable protein kinase A inhibitor PKI 14-22 amide. ELISA showed that while both PGI₂ and NO alone or synergistically raised cAMP, only NO was able to increase intracellular cGMP levels. 3. Treatment of megakaryocytes with PGI₂ abolished both basal and NO-raised cGMP levels. Addition of 8-pCPT-cGMP or activation of soluble guanylyl cyclase by BAY 41-2272 induced cell death in a concentration-dependent manner, and ODQ, an inhibitor of guanylyl cyclase, prevented both PAPA/NO- or BAY 41-2272-induced apoptosis. Specific cGMP phosphodiesterase inhibition by Zaprinast or suppression of adenylyl cyclase by SQ 22536 enhanced the PAPA/NO proapoptotic effect. 4. PGI₂ completely inhibited NO-mediated generation and the increased activity of the cleaved form of caspase-3. 5. In conclusion, our results demonstrate that contrary to their well-known direct and synergistic inhibitory effects on platelets, PGI₂ and NO regulate opposite megakaryocyte survival responses through a delicate balance between intracellular cyclic nucleotide levels and caspase-3 activity control.
format Articulo
Articulo
author Pozner, Roberto Gabriel
Negrotto, Soledad
D'Atri, Lina Paola
Kotler, Mónica Lidia
Lazzari, María Angela
Gómez, Ricardo Martín
Schattner, Mirta
author_facet Pozner, Roberto Gabriel
Negrotto, Soledad
D'Atri, Lina Paola
Kotler, Mónica Lidia
Lazzari, María Angela
Gómez, Ricardo Martín
Schattner, Mirta
author_sort Pozner, Roberto Gabriel
title Prostacyclin prevents nitric oxide-induced megakaryocyte apoptosis
title_short Prostacyclin prevents nitric oxide-induced megakaryocyte apoptosis
title_full Prostacyclin prevents nitric oxide-induced megakaryocyte apoptosis
title_fullStr Prostacyclin prevents nitric oxide-induced megakaryocyte apoptosis
title_full_unstemmed Prostacyclin prevents nitric oxide-induced megakaryocyte apoptosis
title_sort prostacyclin prevents nitric oxide-induced megakaryocyte apoptosis
publishDate 2005
url http://sedici.unlp.edu.ar/handle/10915/127155
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