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Marked Differences in the Effect of Antiepileptic and Cytostatic Drugs on the Functionality of P-Glycoprotein in Human and Rat Brain Capillary Endothelial Cell Lines

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ABSTRACT

Purpose

The expression of P-glycoprotein (Pgp) is increased in brain capillary endothelial cells (BCECs) of patients with pharmacoresistant epilepsy. This may restrict the penetration of antiepileptic drugs (AEDs) into the brain. However, the mechanisms underlying increased Pgp expression in epilepsy patients are not known. One possibility is that AEDs induce the expression and functionality of Pgp in BCECs. Several older AEDs that induce human cytochrome P450 enzymes also induce Pgp in hepatocytes and enterocytes, but whether this extends to Pgp at the human BBB and to newer AEDs is not known.

Methods

This prompted us to study the effects of various old and new AEDs on Pgp functionality in the human BCEC line, hCMEC/D3, using the rhodamine 123 (Rho123) efflux assay. For comparison, experiments were performed in two rat BCEC lines, RBE4 and GPNT, and primary cultures of rat and pig BCECs. Furthermore, known Pgp inducers, such as dexamethasone and several cytostatic drugs, were included in our experiments.

Results

Under control conditions, GPNT cells exhibited the highest and RBE4 the lowest Pgp expression and Rho123 efflux, while intermediate values were determined in hCMEC/D3. Known Pgp inducers increased Rho123 efflux in all cell lines, but marked inter-cell line differences in effect size were observed. Of the various AEDs examined, only carbamazepine (100 μM) moderately increased Pgp functionality in hCMEC/D3, while valproate (300 μM) inhibited Pgp.

Conclusions

These data do not indicate that treatment with AEDs causes a clinically relevant induction in Pgp functionality in BCECs that form the BBB.

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Abbreviations

AEDs:

antiepileptic drugs

ANOVA:

analysis of variance

BBB:

blood–brain barrier

BCEC:

brain capillary endothelial cell

CAR:

constitutive androstane receptor

DMSO:

dimethylsulfoxide

pBCEC:

porcine brain capillary endothelial cell

PBS:

phosphate-buffered saline

Pgp:

P-glycoprotein

PXR:

pregnane X receptor

rBCEC:

rat brain capillary endothelial cell

Rho123:

rhodamine 123

RT:

room temperature

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Acknowledgments and Disclosures

We thank Dr. Pierre-Olivier Couraud (Institut Cochin, Paris, France) for providing hCMEC/D3 cells and Prof. Francoise Roux (INSERM U26, Paris, France) for providing GPNT and RBE4 cells. The study was supported by a grant (Lo 274/10) from the Deutsche Forschungsgemeinschaft (Bonn, Germany). Dana Alms received a PhD scholarship from the FAZIT-Stiftung (Frankfurt am Main, Germany).

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Authors and Affiliations

  1. Department of Pharmacology, Toxicology, and Pharmacy, University of Veterinary Medicine Hannover, Bünteweg 17, 30559, Hannover, Germany

    Dana Alms, Maren Fedrowitz, Kerstin Römermann, Andreas Noack & Wolfgang Löscher

  2. Center for Systems Neuroscience, Hannover, Germany

    Dana Alms & Wolfgang Löscher

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  1. Dana Alms
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  2. Maren Fedrowitz
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Correspondence to Wolfgang Löscher.

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Alms, D., Fedrowitz, M., Römermann, K. et al. Marked Differences in the Effect of Antiepileptic and Cytostatic Drugs on the Functionality of P-Glycoprotein in Human and Rat Brain Capillary Endothelial Cell Lines. Pharm Res 31, 1588–1604 (2014). https://doi.org/10.1007/s11095-013-1264-4

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