Cyclosporin A induced internalization of the bile salt export pump in isolated rat hepatocyte couplets.
Isolated rat hepatocyte couplets were used to perform the comparative study of two widely used immunosuppressors, cyclosporin A (CsA) and tacrolimus (FK506) on hepatocanalicular function. We assessed canalicular function by counting the percentage of couplets that were able to accumulate the fluorescent cholephile, cholyl-lysyl-fluorescein (CLF), into the canalicular vacuole between the two cells, i.e., canalicular vacuole accumulation (CVA) of CLF. Compared to controls (DMSO-treated cells), CsA, in the approximate range of concentrations used therapeutically, caused inhibition of CVA of CLF, disorganization of the bile salt export pump (Bsep) localization at canalicular level resulting in its relocation into the cell, and disruption of the pericanalicular F-actin cytoskeleton. In contrast, FK506, at both approximately therapeutic and supratherapeutic concentrations, had no deleterious effect upon CVA of CLF, upon the localization of the bile salt transporter at the canalicular membrane, or on the organization of the pericanalicular F-actin cytoskeleton. These results point to transporter and cytoskeletal disorganization as contributors or determinants of CsA-induced cholestasis at canalicular level, whereas FK506 does not appear to produce these cholestasis-determining responses even at supratherapeutic concentrations.
Román
Irene D
ID
Departamento de Bioquímica y Biología Molecular, Universidad de Alcalá, E-28871, Alcalá de Henares (Madrid), Spain. lola.roman@uah.es
Fernández-Moreno
M Dolores
MD
Fueyo
Jesús A
JA
Roma
Marcelo G
MG
Coleman
Roger
R
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000818:Animals; D001647:Bile Acids and Salts / Q000378:metabolism; D001648:Bile Canaliculi / Q000187:drug effects* / Q000378:metabolism; D001693:Biological Transport, Active; D002478:Cells, Cultured; D002793:Cholic Acids / Q000378:metabolism; D016572:Cyclosporine / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005452:Fluoresceins / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D007166:Immunosuppressive Agents / Q000633:toxicity*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D016559:Tacrolimus / Q000494:pharmacology
D018528:ATP-Binding Cassette Transporters; D001647:Bile Acids and Salts; D002793:Cholic Acids; D005452:Fluoresceins; D007166:Immunosuppressive Agents; C072313:cholyl-lysylfluorescein; D016572:Cyclosporine; D016559:Tacrolimus