Effect of P-glycoprotein modulators on alkaline phosphatase activity in cultured rat hepatocytes.
Alkaline phosphatases (orthophosphoric-monoester phosphohydrolase, E. C. 3.1.3.1) are a group of nonspecific phosphomonoesterases located primarily in the plasma membrane of the cells in which they occur [1]. It was recently demonstrated that alkaline phosphatase (ALP) concentration in different tissues is positively correlated with the extent of exchange surface per unit volume of the tissue, suggesting an association between ALP and transport systems [2]. Moreover, several groups [3,4,5] obtained evidence of an involvement of ALP in the modulation of P-glycoprotein activity in hepatocytes. The aim of the present study was to determine the putative influence of compounds known to modulate P-glycoprotein-mediated transport on hepatic ALP activity, by using primary cultured rat hepatocytes. The K(m) and V(max) values of ALP were determined (657.2 microM (306.8-933.1) and 32.0+/-1.5 nmol mg protein(-1) min(-1), respectively). Vanadate and corticosterone concentration-dependently reduced ALP activity, producing maximal reductions of 79% (100 microM) and 71% (100 microM), respectively. The IC50's were found to be 7.9 microM (2.1-29.5 microM) and 2.4 microM (0.2-35.2 microM), respectively. Cyclosporin A, verapamil, octreotide, kaempferol, daunomycin and genistein produced a concentration-dependent increase in ALP activity. ALP activity was maximally increased to 253%, 390%, 180%, 487%, 449% and 193% of control in the presence of 100 microM cyclosporin A, 50 microM verapamil, 10 microM octreotide, 100 microM kaempferol, 100 microM daunomycin and 1 microM genistein, respectively. The results show that all P-glycoprotein modulators tested were able to significantly affect the activity of hepatic-ALP. These effects on ALP activity may contribute to the modulation of P-glycoprotein activity by these drugs.
Calhau
C
C
Department of Biochemistry, Faculty of Medicine, 4200 Porto, Portugal. ccalhau@med.up.pt
Martel
F
F
Hipólito-Reis
C
C
Azevedo
I
I
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000187:drug effects / Q000378:metabolism*; D000305:Adrenal Cortex Hormones / Q000494:pharmacology; D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D007700:Kinetics; D007978:Levamisole / Q000494:pharmacology; D008297:Male; D009596:Nitrophenols / Q000378:metabolism; D015282:Octreotide / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D014638:Vanadates / Q000494:pharmacology
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000305:Adrenal Cortex Hormones; D004791:Enzyme Inhibitors; D009596:Nitrophenols; D007978:Levamisole; D014638:Vanadates; D000469:Alkaline Phosphatase; D015282:Octreotide; C024836:4-nitrophenol