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A Method to Assess Functional Activity of P-Glycoprotein in Vitro Based on the Energy Requirements of the Transporter

  • A. Reichel
  • Z.-D. Reeve-Chen
  • D. J. Begley
  • N. J. Abbott
Part of the Advances in Behavioral Biology book series (ABBI, volume 46)

Summary

P-glycoprotein (Pgp) is constitutively expressed in the luminal membrane of endothelial cells forming the blood-brain barrier. Pgp acts as an active drug efflux pump and hence frustrates treatment of brain diseases by preventing a large variety of chemically unrelated drugs from entering the central nervous system. The screening for non-toxic, specific inhibitors requires a large number of compounds to be tested for their possible interaction with Pgp. As brain endothelial cells retain expression of Pgp in culture, in vitro assays are the obvious choice. The quality of such a screening model relys on a maintained expression of Pgp which needs to be characterised beforehand. Thus, we designed a simple protocol based on energy requirements of Pgp mediated drug efflux to assess the functional activity of the transporter quantitatively.

Keywords

Brain Endothelial Cell Cellular Accumulation Active Efflux RBE4 Cell Brain Microvessel Endothelial Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Résumé

La P-glycoprotéine (P-gp) est exprimée constitutionnellement au niveau de la membrane luminale des cellules endothéliales qui forment la barrière hémato-encéphalique. La P-gp agit comme une pompe active pour expulser les médicaments et donc empêche le traitement des maladies cérébrales, en empêchant un certain nombre de médicaments, de structures chimiques diverses, de pénétrer dans le système nerveux central. Il faut donc tester un grand nombre de composés par rapport à leur interaction possible avec la P-gp, pour définir des inhibiteurs spécifiques non toxiques.Comrne les cellules endothéliales cérébrales en culture expriment encore la P-gp, il est judicieux de faire des tests in vitro. La qualité d’un tel critère de choix repose sur cette expression maintenue de la P-gp, qui doit être vérifiée. Nous avons donc défini un protocole simple basé sur les besoins énergétiques de la P-gp pour l’efflux des drogues pour contrôler quantitativement l’activité fonctionnelle du transporteur.

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Copyright information

© Springer Science+Business Media New York 1996

Authors and Affiliations

  • A. Reichel
    • 1
  • Z.-D. Reeve-Chen
    • 1
  • D. J. Begley
    • 1
  • N. J. Abbott
    • 1
  1. 1.Physiology Group Biomedical Sciences DivisionKing’s College LondonLondonUK

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