Inward transport of [3H]-1-methyl-4-phenylpyridinium in rat isolated hepatocytes: putative involvement of a P-glycoprotein transporter.
1. The liver has an important role in the detoxification of organic cations from the circulation. [3H]-1-methyl-4-phenylpyridinium ([3H]-MPP+), a low molecular weight organic cation, is efficiently taken up and accumulated by rat hepatocytes through mechanisms partially unknown. 2. The aim of the present work was to characterize further the uptake of MPP+ by rat isolated hepatocytes. The putative interactions of a wide range of drugs, including inhibitors/substrates of P-glycoprotein, were studied. 3. The uptake of MPP+ was investigated in rat freshly isolated hepatocytes (incubated in Krebs-Henseleit medium with 200 nM [3H]-MPP+ for 5 min) and in the rat liver in situ (perfused with Krebs-Henseleit/BSA medium with 200 nM [3H]-MPP+ for 30 min). [3H]-MPP+ accumulation in the cells and in tissue was determined by liquid scintillation counting. 4. Verapamil (100 microM), quinidine (100 microM), amiloride (1 mM), (+)-tubocurarine (100 microM), vecuronium (45 microM), bilirubin (200 microM), progesterone (200 microM), daunomycin (100 microM), vinblastine (100 microM), cyclosporin A (100 microM) and cimetidine (100 microM) had a significant inhibitory effect on the accumulation of [3H]-MPP+ in isolated hepatocytes. Tetraethylammonium (100 microM) had no effect. 5. In the rat perfused liver, both cyclosporin A (100 microM) and verapamil (100 microM) had much less marked inhibitory effects as compared to their effects on isolated hepatocytes (0% against 35% and 45% against 96% of inhibition, respectively). 6. Inhibition of alkaline phosphatase activity by increasing or decreasing the pH of the incubation medium or by the presence of vanadate (1 mM) or homoarginine (500 microM) led to a significant increase in the accumulation of [3H]-MPP+ in isolated hepatocytes. 7. It was concluded that, in addition to the type I organic cation hepatic transporter, [3H]-MPP+ is taken up by rat hepatocytes through P-glycoprotein, a canalicular transport system that usually excretes endobiotics and xenobiotics. We proposed that the reversal of transport through P-glycoprotein may be related to the loss of efficacy of alkaline in isolated hepatocytes.
Martel
F
F
Department of Biochemistry, Faculty of Medicine, Porto, Portugal.
Martins
M J
MJ
Hipólito-Reis
C
C
Azevedo
I
I
D015655:1-Methyl-4-phenylpyridinium / Q000378:metabolism*; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism*; D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors / Q000378:metabolism; D000818:Animals; D000963:Antimetabolites / Q000494:pharmacology; D003470:Culture Media; D015259:Dopamine Agents / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007473:Ion Channels / Q000187:drug effects / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D008297:Male; D051381:Rats; D017208:Rats, Wistar
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000963:Antimetabolites; D003470:Culture Media; D015259:Dopamine Agents; D007473:Ion Channels; D000469:Alkaline Phosphatase; D015655:1-Methyl-4-phenylpyridinium