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Connexin 43 and metabolic effect of fatty acids in stressed endothelial cells
Changes in the inner mitochondrial membrane potential (∆ψ) may lead either to apoptosis or to protective autophagy. Connexin 43 (Cx43), a gap junction protein, is suggested to affect mitochondrial membrane permeability. The aim of our study was to analyze Cx43 gene expression, Cx43 protein localizat...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer-Verlag
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3316752/ https://www.ncbi.nlm.nih.gov/pubmed/21948354 http://dx.doi.org/10.1007/s12263-011-0247-5 |
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author | Kiec-Wilk, Beata Czech, Urszula Janczarska, Katarzyna Knapp, Anna Goralska, Joanna Cialowicz, Urszula Malecki, Maciej T. Dembinska-Kiec, Aldona |
author_facet | Kiec-Wilk, Beata Czech, Urszula Janczarska, Katarzyna Knapp, Anna Goralska, Joanna Cialowicz, Urszula Malecki, Maciej T. Dembinska-Kiec, Aldona |
author_sort | Kiec-Wilk, Beata |
collection | PubMed |
description | Changes in the inner mitochondrial membrane potential (∆ψ) may lead either to apoptosis or to protective autophagy. Connexin 43 (Cx43), a gap junction protein, is suggested to affect mitochondrial membrane permeability. The aim of our study was to analyze Cx43 gene expression, Cx43 protein localization and mitochondrial function in the human endothelial cells stressed by dietary-free fatty acids (FFA) and TNFα. Human endothelial cells (HUVECs) were incubated with (10–30 uM) palmitic (PA), oleic (OA), eicosapentaenoic (EPA) or arachidonic (AA) acids for 24 h. TNFα (5 ng/ml) was added at the last 4 h of incubation. The Cx43 gene expression was analyzed by the quantitative real-time PCR. The Cx43 protein concentrations in whole cells and in the isolated mitochondria were measured. Changes in ∆ψ and Cx43 localization were analyzed by flow cytometry or fluorescence microscopy. Generated ATP was measured by a luminescence assay. TNFα, PA and OA significantly decreased ∆ψ, while AA (P = 0.047) and EPA (P = 0.004) increased ∆ψ value. Preincubation with EPA or AA partially prevented the TNFα-induced decrease of ∆ψ. Incubation with AA resulted in up-regulation of the Cx43 gene expression. AA or PA significantly increased Cx43 protein content; however, presence of TNFα in general aggravated the negative effect of FFA. Only EPA was found to increase ATP generation in HUVECs. The fatty acid-specific induction of changes in Cx43 expression and protein concentration as well as the normalization of ∆ψ and increase of ATP generation seem to be the separate, independent mechanisms of FFA-mediated modulatory effect in the human endothelial cells pathology. |
format | Online Article Text |
id | pubmed-3316752 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Springer-Verlag |
record_format | MEDLINE/PubMed |
spelling | pubmed-33167522012-04-03 Connexin 43 and metabolic effect of fatty acids in stressed endothelial cells Kiec-Wilk, Beata Czech, Urszula Janczarska, Katarzyna Knapp, Anna Goralska, Joanna Cialowicz, Urszula Malecki, Maciej T. Dembinska-Kiec, Aldona Genes Nutr Research Paper Changes in the inner mitochondrial membrane potential (∆ψ) may lead either to apoptosis or to protective autophagy. Connexin 43 (Cx43), a gap junction protein, is suggested to affect mitochondrial membrane permeability. The aim of our study was to analyze Cx43 gene expression, Cx43 protein localization and mitochondrial function in the human endothelial cells stressed by dietary-free fatty acids (FFA) and TNFα. Human endothelial cells (HUVECs) were incubated with (10–30 uM) palmitic (PA), oleic (OA), eicosapentaenoic (EPA) or arachidonic (AA) acids for 24 h. TNFα (5 ng/ml) was added at the last 4 h of incubation. The Cx43 gene expression was analyzed by the quantitative real-time PCR. The Cx43 protein concentrations in whole cells and in the isolated mitochondria were measured. Changes in ∆ψ and Cx43 localization were analyzed by flow cytometry or fluorescence microscopy. Generated ATP was measured by a luminescence assay. TNFα, PA and OA significantly decreased ∆ψ, while AA (P = 0.047) and EPA (P = 0.004) increased ∆ψ value. Preincubation with EPA or AA partially prevented the TNFα-induced decrease of ∆ψ. Incubation with AA resulted in up-regulation of the Cx43 gene expression. AA or PA significantly increased Cx43 protein content; however, presence of TNFα in general aggravated the negative effect of FFA. Only EPA was found to increase ATP generation in HUVECs. The fatty acid-specific induction of changes in Cx43 expression and protein concentration as well as the normalization of ∆ψ and increase of ATP generation seem to be the separate, independent mechanisms of FFA-mediated modulatory effect in the human endothelial cells pathology. Springer-Verlag 2011-09-24 /pmc/articles/PMC3316752/ /pubmed/21948354 http://dx.doi.org/10.1007/s12263-011-0247-5 Text en © The Author(s) 2011 https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited. |
spellingShingle | Research Paper Kiec-Wilk, Beata Czech, Urszula Janczarska, Katarzyna Knapp, Anna Goralska, Joanna Cialowicz, Urszula Malecki, Maciej T. Dembinska-Kiec, Aldona Connexin 43 and metabolic effect of fatty acids in stressed endothelial cells |
title | Connexin 43 and metabolic effect of fatty acids in stressed endothelial cells |
title_full | Connexin 43 and metabolic effect of fatty acids in stressed endothelial cells |
title_fullStr | Connexin 43 and metabolic effect of fatty acids in stressed endothelial cells |
title_full_unstemmed | Connexin 43 and metabolic effect of fatty acids in stressed endothelial cells |
title_short | Connexin 43 and metabolic effect of fatty acids in stressed endothelial cells |
title_sort | connexin 43 and metabolic effect of fatty acids in stressed endothelial cells |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3316752/ https://www.ncbi.nlm.nih.gov/pubmed/21948354 http://dx.doi.org/10.1007/s12263-011-0247-5 |
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