To test the activity of P-170 glycoprotein in isolated hepatocytes, a method has been developed employing the fluorescent dye rhodamine 123 (R-123). The uptake of R-123 by both freshly isolated and 4-hr-plated hepatocytes depends on dye concentration, time of incubation, and cell number. The efflux of R-123 from cells is inhibited by sodium azide and by verapamil. In standard conditions the efficiency of efflux of R-123 from cells correlates with the relative amount of immunoblottable glycoprotein. The method has been applied to detection of P-170 activity in hepatocytes from animals of different ages as well as from carcinogen-treated animals. The proposed assay appears a simple and adequate tool for the functional assessment of multidrug transporter in liver.
|Autori interni:||PAOLICCHI, ALDO|
|Autori:||Chieli E; Santoni Rugiu E; Cervelli F; Paolicchi A; Tongiani R|
|Titolo:||Assessment of P-glycoprotein-dependent drug transport in isolated rat hepatocytes using rhodamine 123.|
|Anno del prodotto:||1993|
|Appare nelle tipologie:||1.1 Articolo in rivista|