Lindner, Christian and Sigrüner, Alexander and Walther, Franziska and Bogdahn, Ulrich and Couraud, Pierre O. and Schmitz, Gerd and Schlachetzki, Felix (2012) ATP-binding cassette transporters in immortalised human brain microvascular endothelial cells in normal and hypoxic conditions. Experimental & Translational Stroke Medicine 4, p. 9.
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PDF - Published Version
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.
We exposed hCMEC/D3 cells to 24 hours of hypoxia alone and to hypoxia followed by 60min of reoxygenisation as a in vitro model for I/R. Western blot showed mild upregulation of hypoxia inducible factor (HIF-1alpha) 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.
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.
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 including neuronal stem cells inducing an overall ischemic response at the BBB.
|Date:||3 May 2012|
|Institutions:|| Medicine > Lehrstuhl für Klinische Chemie und Laboratoriumsmedizin|
Medicine > Lehrstuhl für Neurologie
Medicine > Lehrstuhl für Psychiatrie und Psychotherapie
|Projects:||Open Access Publizieren (DFG)|
|Keywords:||Blood–brain barrier; Ischemia/reperfusion injury; Hypoxia-inducible factor; Multidrug resistance; ABC transporters; Stroke|
|Subjects:||600 Technology > 610 Medical sciences Medicine|
|Refereed:||Yes, this version has been refereed|
|Created at the University of Regensburg:||Partially|
|Deposited On:||07 Sep 2012 06:48|
|Last Modified:||21 Mar 2013 09:31|