Characterization of metformin transport system in NIH 3T3 cells.
The biochemical properties of the metformin transport system were studied in NIH 3T3 cells. 14C-metformin uptake appeared to be a sodium dependent process. Iso-osmotical replacement of Na+ by choline chloride in the assay medium resulted in a decrease of metformin uptake. Amiloride (200 microM) inhibited the metformin transport by 35% in these cells. Gramicidin, a channel ionophore, was the most effective in inhibiting the metformin transport as compared to valinomycin, a mobile ion carrier, and Ca2+ ionophore (A 23187). Loading of cells with asparagine, ornithine, or polylysine did not influence the uptake process. However, the addition of lysine or arginine significantly stimulated the metformin uptake by NIH 3T3 cells. Similarly, the addition of metformin stimulated the arginine uptake by these cells, suggesting that metformin shares the y+ transport system. Metformin inhibited competitively the uptake of 14C-spermidine, a molecule of the polyamine family, by NIH 3T3 cells, whereas the latter failed to influence the uptake of the former significantly by these cells. Incubation of NIH 3T3 cells in the presence of difluoromethyl-ornithine (a suicidal inhibitor of polyamine biosynthesis) stimulated the spermidine, but not the metformin, uptake by these cells. Interestingly, a prolonged incubation of these cells in the presence of metformin failed to down-regulate the spermidine transport process. The spermidine- and methylglyoxal-bis(guanylhydrazone), MGBG-transport deficient (3T3MG) cells which do not accumulate exogeneous spermidine or MGBG, took up 14C-metformin. However, 14C-metformin uptake by 3T3MG cells was lower than that by normal NIH 3T3 cells.
Khan
N A
NA
Unité Fonctionnelle de Biologie Cellulaire, Faculté de Médecine, Université de Rennes, France.
Wiernsperger
N
N
Quemener
V
V
Havouis
R
R
Moulinoux
J P
JP
D016475:3T3 Cells / Q000378:metabolism*; D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D005110:Extracellular Space / Q000378:metabolism; D007476:Ionophores / Q000494:pharmacology; D008687:Metformin / Q000493:pharmacokinetics* / Q000494:pharmacology; D051379:Mice; D008935:Mitoguazone / Q000494:pharmacology; D009154:Mutation; D011073:Polyamines / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D013095:Spermidine / Q000493:pharmacokinetics / Q000494:pharmacology; D013997:Time Factors
D000596:Amino Acids; D007476:Ionophores; D011073:Polyamines; D008687:Metformin; D012964:Sodium; D008935:Mitoguazone; D013095:Spermidine