Cargando…
The Association of Ascorbic Acid, Deferoxamine and N-Acetylcysteine Improves Cardiac Fibroblast Viability and Cellular Function Associated with Tissue Repair Damaged by Simulated Ischemia/Reperfusion
Acute myocardial infarction is one of the leading causes of death worldwide and thus, an extensively studied disease. Nonetheless, the effects of ischemia/reperfusion injury elicited by oxidative stress on cardiac fibroblast function associated with tissue repair are not completely understood. Ascor...
Autores principales: | , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6943610/ https://www.ncbi.nlm.nih.gov/pubmed/31817022 http://dx.doi.org/10.3390/antiox8120614 |
_version_ | 1783484914975375360 |
---|---|
author | Parra-Flores, Pablo Riquelme, Jaime A Valenzuela-Bustamante, Paula Leiva-Navarrete, Sebastian Vivar, Raúl Cayupi-Vivanco, Jossete Castro, Esteban Espinoza-Pérez, Claudio Ruz-Cortés, Felipe Pedrozo, Zully Lavandero, Sergio Rodrigo, Ramon Diaz-Araya, Guillermo |
author_facet | Parra-Flores, Pablo Riquelme, Jaime A Valenzuela-Bustamante, Paula Leiva-Navarrete, Sebastian Vivar, Raúl Cayupi-Vivanco, Jossete Castro, Esteban Espinoza-Pérez, Claudio Ruz-Cortés, Felipe Pedrozo, Zully Lavandero, Sergio Rodrigo, Ramon Diaz-Araya, Guillermo |
author_sort | Parra-Flores, Pablo |
collection | PubMed |
description | Acute myocardial infarction is one of the leading causes of death worldwide and thus, an extensively studied disease. Nonetheless, the effects of ischemia/reperfusion injury elicited by oxidative stress on cardiac fibroblast function associated with tissue repair are not completely understood. Ascorbic acid, deferoxamine, and N-acetylcysteine (A/D/N) are antioxidants with known cardioprotective effects, but the potential beneficial effects of combining these antioxidants in the tissue repair properties of cardiac fibroblasts remain unknown. Thus, the aim of this study was to evaluate whether the pharmacological association of these antioxidants, at low concentrations, could confer protection to cardiac fibroblasts against simulated ischemia/reperfusion injury. To test this, neonatal rat cardiac fibroblasts were subjected to simulated ischemia/reperfusion in the presence or absence of A/D/N treatment added at the beginning of simulated reperfusion. Cell viability was assessed using trypan blue staining, and intracellular reactive oxygen species (ROS) production was assessed using a 2′,7′-dichlorofluorescin diacetate probe. Cell death was measured by flow cytometry using propidium iodide. Cell signaling mechanisms, differentiation into myofibroblasts and pro-collagen I production were determined by Western blot, whereas migration was evaluated using the wound healing assay. Our results show that A/D/N association using a low concentration of each antioxidant increased cardiac fibroblast viability, but that their separate administration did not provide protection. In addition, A/D/N association attenuated oxidative stress triggered by simulated ischemia/reperfusion, induced phosphorylation of pro-survival extracellular-signal-regulated kinases 1/2 (ERK1/2) and PKB (protein kinase B)/Akt, and decreased phosphorylation of the pro-apoptotic proteins p38- mitogen-activated protein kinase (p38-MAPK) and c-Jun-N-terminal kinase (JNK). Moreover, treatment with A/D/N also reduced reperfusion-induced apoptosis, evidenced by a decrease in the sub-G1 population, lower fragmentation of pro-caspases 9 and 3, as well as increased B-cell lymphoma-extra large protein (Bcl-xL)/Bcl-2-associated X protein (Bax) ratio. Furthermore, simulated ischemia/reperfusion abolished serum-induced migration, TGF-β1 (transforming growth factor beta 1)-mediated cardiac fibroblast-to-cardiac myofibroblast differentiation, and angiotensin II-induced pro-collagen I synthesis, but these effects were prevented by treatment with A/D/N. In conclusion, this is the first study where a pharmacological combination of A/D/N, at low concentrations, protected cardiac fibroblast viability and function after simulated ischemia/reperfusion, and thereby represents a novel therapeutic approach for cardioprotection. |
format | Online Article Text |
id | pubmed-6943610 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-69436102020-01-10 The Association of Ascorbic Acid, Deferoxamine and N-Acetylcysteine Improves Cardiac Fibroblast Viability and Cellular Function Associated with Tissue Repair Damaged by Simulated Ischemia/Reperfusion Parra-Flores, Pablo Riquelme, Jaime A Valenzuela-Bustamante, Paula Leiva-Navarrete, Sebastian Vivar, Raúl Cayupi-Vivanco, Jossete Castro, Esteban Espinoza-Pérez, Claudio Ruz-Cortés, Felipe Pedrozo, Zully Lavandero, Sergio Rodrigo, Ramon Diaz-Araya, Guillermo Antioxidants (Basel) Article Acute myocardial infarction is one of the leading causes of death worldwide and thus, an extensively studied disease. Nonetheless, the effects of ischemia/reperfusion injury elicited by oxidative stress on cardiac fibroblast function associated with tissue repair are not completely understood. Ascorbic acid, deferoxamine, and N-acetylcysteine (A/D/N) are antioxidants with known cardioprotective effects, but the potential beneficial effects of combining these antioxidants in the tissue repair properties of cardiac fibroblasts remain unknown. Thus, the aim of this study was to evaluate whether the pharmacological association of these antioxidants, at low concentrations, could confer protection to cardiac fibroblasts against simulated ischemia/reperfusion injury. To test this, neonatal rat cardiac fibroblasts were subjected to simulated ischemia/reperfusion in the presence or absence of A/D/N treatment added at the beginning of simulated reperfusion. Cell viability was assessed using trypan blue staining, and intracellular reactive oxygen species (ROS) production was assessed using a 2′,7′-dichlorofluorescin diacetate probe. Cell death was measured by flow cytometry using propidium iodide. Cell signaling mechanisms, differentiation into myofibroblasts and pro-collagen I production were determined by Western blot, whereas migration was evaluated using the wound healing assay. Our results show that A/D/N association using a low concentration of each antioxidant increased cardiac fibroblast viability, but that their separate administration did not provide protection. In addition, A/D/N association attenuated oxidative stress triggered by simulated ischemia/reperfusion, induced phosphorylation of pro-survival extracellular-signal-regulated kinases 1/2 (ERK1/2) and PKB (protein kinase B)/Akt, and decreased phosphorylation of the pro-apoptotic proteins p38- mitogen-activated protein kinase (p38-MAPK) and c-Jun-N-terminal kinase (JNK). Moreover, treatment with A/D/N also reduced reperfusion-induced apoptosis, evidenced by a decrease in the sub-G1 population, lower fragmentation of pro-caspases 9 and 3, as well as increased B-cell lymphoma-extra large protein (Bcl-xL)/Bcl-2-associated X protein (Bax) ratio. Furthermore, simulated ischemia/reperfusion abolished serum-induced migration, TGF-β1 (transforming growth factor beta 1)-mediated cardiac fibroblast-to-cardiac myofibroblast differentiation, and angiotensin II-induced pro-collagen I synthesis, but these effects were prevented by treatment with A/D/N. In conclusion, this is the first study where a pharmacological combination of A/D/N, at low concentrations, protected cardiac fibroblast viability and function after simulated ischemia/reperfusion, and thereby represents a novel therapeutic approach for cardioprotection. MDPI 2019-12-03 /pmc/articles/PMC6943610/ /pubmed/31817022 http://dx.doi.org/10.3390/antiox8120614 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Parra-Flores, Pablo Riquelme, Jaime A Valenzuela-Bustamante, Paula Leiva-Navarrete, Sebastian Vivar, Raúl Cayupi-Vivanco, Jossete Castro, Esteban Espinoza-Pérez, Claudio Ruz-Cortés, Felipe Pedrozo, Zully Lavandero, Sergio Rodrigo, Ramon Diaz-Araya, Guillermo The Association of Ascorbic Acid, Deferoxamine and N-Acetylcysteine Improves Cardiac Fibroblast Viability and Cellular Function Associated with Tissue Repair Damaged by Simulated Ischemia/Reperfusion |
title | The Association of Ascorbic Acid, Deferoxamine and N-Acetylcysteine Improves Cardiac Fibroblast Viability and Cellular Function Associated with Tissue Repair Damaged by Simulated Ischemia/Reperfusion |
title_full | The Association of Ascorbic Acid, Deferoxamine and N-Acetylcysteine Improves Cardiac Fibroblast Viability and Cellular Function Associated with Tissue Repair Damaged by Simulated Ischemia/Reperfusion |
title_fullStr | The Association of Ascorbic Acid, Deferoxamine and N-Acetylcysteine Improves Cardiac Fibroblast Viability and Cellular Function Associated with Tissue Repair Damaged by Simulated Ischemia/Reperfusion |
title_full_unstemmed | The Association of Ascorbic Acid, Deferoxamine and N-Acetylcysteine Improves Cardiac Fibroblast Viability and Cellular Function Associated with Tissue Repair Damaged by Simulated Ischemia/Reperfusion |
title_short | The Association of Ascorbic Acid, Deferoxamine and N-Acetylcysteine Improves Cardiac Fibroblast Viability and Cellular Function Associated with Tissue Repair Damaged by Simulated Ischemia/Reperfusion |
title_sort | association of ascorbic acid, deferoxamine and n-acetylcysteine improves cardiac fibroblast viability and cellular function associated with tissue repair damaged by simulated ischemia/reperfusion |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6943610/ https://www.ncbi.nlm.nih.gov/pubmed/31817022 http://dx.doi.org/10.3390/antiox8120614 |
work_keys_str_mv | AT parraflorespablo theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT riquelmejaimea theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT valenzuelabustamantepaula theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT leivanavarretesebastian theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT vivarraul theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT cayupivivancojossete theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT castroesteban theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT espinozaperezclaudio theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT ruzcortesfelipe theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT pedrozozully theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT lavanderosergio theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT rodrigoramon theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT diazarayaguillermo theassociationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT parraflorespablo associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT riquelmejaimea associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT valenzuelabustamantepaula associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT leivanavarretesebastian associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT vivarraul associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT cayupivivancojossete associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT castroesteban associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT espinozaperezclaudio associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT ruzcortesfelipe associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT pedrozozully associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT lavanderosergio associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT rodrigoramon associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion AT diazarayaguillermo associationofascorbicaciddeferoxamineandnacetylcysteineimprovescardiacfibroblastviabilityandcellularfunctionassociatedwithtissuerepairdamagedbysimulatedischemiareperfusion |