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ATP-binding cassette transporters in immortalised human brain microvascular endothelial cells in normal and hypoxic conditions

BACKGROUND: Rapid reperfusion following ischemia is the most effective therapy in stroke therapy. However, the success may be compromised by ischemia & reperfusion (I/R) injury and at the human blood–brain barrier (BBB), therefore the effects on transendothelial transport are of special interest...

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Autores principales: Lindner, Christian, Sigrüner, Alexander, Walther, Franziska, Bogdahn, Ulrich, Couraud, Pierre O, Schmitz, Gert, Schlachetzki, Felix
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3466144/
https://www.ncbi.nlm.nih.gov/pubmed/22553972
http://dx.doi.org/10.1186/2040-7378-4-9
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author Lindner, Christian
Sigrüner, Alexander
Walther, Franziska
Bogdahn, Ulrich
Couraud, Pierre O
Schmitz, Gert
Schlachetzki, Felix
author_facet Lindner, Christian
Sigrüner, Alexander
Walther, Franziska
Bogdahn, Ulrich
Couraud, Pierre O
Schmitz, Gert
Schlachetzki, Felix
author_sort Lindner, Christian
collection PubMed
description BACKGROUND: Rapid reperfusion following ischemia is the most effective therapy in stroke therapy. However, the success may be compromised by ischemia & reperfusion (I/R) injury and at the human blood–brain barrier (BBB), therefore the effects on transendothelial transport are of special interest. Current studies suggest the ATP-binding cassette (ABC) transporters to be regulated upon ischemic stroke in a way that impedes the effects of drug therapy. The immortalised human brain microvascular endothelial cell line hCMEC/D3 provides most of the unique properties of the BBB with respect to transport and might be a reliable in vitro model to study transendothelial transport after I/R. METHODS: We exposed hCMEC/D3 cells to 24 hours of hypoxia alone and to hypoxia followed by 60 min of reoxygenisation as an in vitro model for I/R. Western blot showed mild upregulation of hypoxia inducible factor (HIF-1α) after hypoxia alone and RNA lysates were analysed with a well-established real-time RT-PCR-based TaqMan low-density array detecting 47 of 48 known human ABC transporters. RESULTS: No significant increases of ABC mRNA expression levels were detected neither in hypoxic nor in I/R samples. However, slight decrease of ABCC1 in hypoxic and I/R samples and of ABCA10 and ABCD3 in I/R samples was observed. CONCLUSION: Our data suggests that hCMEC/D3 cell line and – at the moment – in vitro models in general are a poor basis for stroke research but may be enhanced by co-culturing more cells of the neurovascular unit inducing an overall ischemic response at the BBB.
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spelling pubmed-34661442012-10-09 ATP-binding cassette transporters in immortalised human brain microvascular endothelial cells in normal and hypoxic conditions Lindner, Christian Sigrüner, Alexander Walther, Franziska Bogdahn, Ulrich Couraud, Pierre O Schmitz, Gert Schlachetzki, Felix Exp Transl Stroke Med Research BACKGROUND: Rapid reperfusion following ischemia is the most effective therapy in stroke therapy. However, the success may be compromised by ischemia & reperfusion (I/R) injury and at the human blood–brain barrier (BBB), therefore the effects on transendothelial transport are of special interest. Current studies suggest the ATP-binding cassette (ABC) transporters to be regulated upon ischemic stroke in a way that impedes the effects of drug therapy. The immortalised human brain microvascular endothelial cell line hCMEC/D3 provides most of the unique properties of the BBB with respect to transport and might be a reliable in vitro model to study transendothelial transport after I/R. METHODS: We exposed hCMEC/D3 cells to 24 hours of hypoxia alone and to hypoxia followed by 60 min of reoxygenisation as an in vitro model for I/R. Western blot showed mild upregulation of hypoxia inducible factor (HIF-1α) after hypoxia alone and RNA lysates were analysed with a well-established real-time RT-PCR-based TaqMan low-density array detecting 47 of 48 known human ABC transporters. RESULTS: No significant increases of ABC mRNA expression levels were detected neither in hypoxic nor in I/R samples. However, slight decrease of ABCC1 in hypoxic and I/R samples and of ABCA10 and ABCD3 in I/R samples was observed. CONCLUSION: Our data suggests that hCMEC/D3 cell line and – at the moment – in vitro models in general are a poor basis for stroke research but may be enhanced by co-culturing more cells of the neurovascular unit inducing an overall ischemic response at the BBB. BioMed Central 2012-05-03 /pmc/articles/PMC3466144/ /pubmed/22553972 http://dx.doi.org/10.1186/2040-7378-4-9 Text en Copyright ©2012 Lindner et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Lindner, Christian
Sigrüner, Alexander
Walther, Franziska
Bogdahn, Ulrich
Couraud, Pierre O
Schmitz, Gert
Schlachetzki, Felix
ATP-binding cassette transporters in immortalised human brain microvascular endothelial cells in normal and hypoxic conditions
title ATP-binding cassette transporters in immortalised human brain microvascular endothelial cells in normal and hypoxic conditions
title_full ATP-binding cassette transporters in immortalised human brain microvascular endothelial cells in normal and hypoxic conditions
title_fullStr ATP-binding cassette transporters in immortalised human brain microvascular endothelial cells in normal and hypoxic conditions
title_full_unstemmed ATP-binding cassette transporters in immortalised human brain microvascular endothelial cells in normal and hypoxic conditions
title_short ATP-binding cassette transporters in immortalised human brain microvascular endothelial cells in normal and hypoxic conditions
title_sort atp-binding cassette transporters in immortalised human brain microvascular endothelial cells in normal and hypoxic conditions
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3466144/
https://www.ncbi.nlm.nih.gov/pubmed/22553972
http://dx.doi.org/10.1186/2040-7378-4-9
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