Molecular and Cellular PharmacologyP-glycoprotein-mediated transport of morphine in brain capillary endothelial cells
Section snippets
Materials
Morphine sulfate, verapamil hydrochloride, CyA, and sodium azide were purchased from the Sigma Chemical Co. Rhodamine 123 was purchased from Molecular Probes. GF120918 was donated by Glaxo Wellcome, Inc.
[3H]Morphine (84.5 Ci/mmol) was purchased from New England Nuclear Life Sciences Products and was > 98.5% pure as determined by HPLC. [3H]Propranolol (15 Ci/mmol) and [14C]sucrose (0.442 Ci/mmol) were purchased from Amersham International plc. All drugs were used without further purification.
All
Characterization of cell culture model
Bovine brain capillary endothelial cells were isolated and cultured successfully. Cells grew to a confluent monolayer within 14 days and were used for cellular accumulation, transendothelial permeability, and efflux studies on days 14–15. An electron micrograph of the BBCEC monolayer grown on the Transwell™ polycarbonate membrane demonstrates the presence of a monolayer of cells and junctional complexes between the endothelial cells (Fig. 1). The cells did not appear to migrate through the
Discussion
Brain capillary endothelial cells are the primary physical barrier between brain extracellular fluid and blood. Passage of morphine through the BBB is well recognized, but compared with many drugs the penetration is limited. Early studies have demonstrated that the brain uptake of morphine in rats after carotid injection is very low 18, 19. Observed morphine brain-to-plasma concentration ratios of less than unity provide additional evidence of the limited BBB permeability of morphine 20, 21, 22
Acknowledgements
This work was supported, in part, by an American Foundation for Pharmaceutical Education Pre-Doctoral Fellowship (S. P. Letrent). This study was presented, in part, at the Eleventh Annual Meeting of the American Association of Pharmaceutical Scientists, November 2–6, 1997, Boston, Massachusetts. The assistance of J. Lee (Glaxo Wellcome, Inc.) for electron micrograph work is gratefully acknowledged.
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