Hepatocellular uptake of aflatoxin B1 by non-ionic diffusion. Inhibition of bile acid transport by interference with membrane lipids.
Aflatoxin B1 permeates isolated rat hepatocytes by non-ionic diffusion. Its uptake is neither saturable nor influenced by metabolic energy and not inhibited by treatment of cells with proteases. The initial rate of aflatoxin B1 uptake measured at 7 degrees C is between 40 and 50% compared to that at 37 degrees C. However, after an incubation period of 7 minutes identical equilibrium uptake is reached at both temperatures. The apparent activation energies, calculated for aflatoxin B1 uptake by Arrhenius diagrams ranged between 1.69 and 4.5 kcal/mol. A Q10 value of 1.34 was calculated for a temperature interval of 7-17 degrees C but decreased to 1.05 for the interval of 27-37 degrees C. Liposomes or lipoproteins added to the cell suspension inhibited the aflatoxin B1 uptake into hepatocytes. Liposomes mainly composed of unsaturated fatty acids bind twice as much aflatoxin B1 as those composed of saturated ones, indicating that the lipophilicity of the mycotoxin is crucial in the determination of its uptake into liver cells. At concentrations above 5 micrograms/ml, aflatoxin B1 inhibited the carrier-mediated uptake of cholic acid and of phalloidin into hepatocytes. This effect was reversible and abolished by washing the cells after preincubation with aflatoxin. In concentrations below 5 micrograms/ml the uptake of phallotoxin and cholic acid was however stimulated by 15-25%. These results indicate, that a carrier-mediated uptake into hepatocytes via the multispecific bile salt transporter is not responsible for the organoselective clearance of aflatoxins by the liver. On the other hand, the cholestatic effect of aflatoxin B1 results at least partially from the inhibition of the multispecific bile acid transport system. This inhibition may arise from affinity of aflatoxins to lipid domains of the cell membrane.
D016604:Aflatoxin B1; D000348:Aflatoxins / Q000493:pharmacokinetics* / Q000494:pharmacology; D000818:Animals; D001647:Bile Acids and Salts / Q000378:metabolism*; D001692:Biological Transport / Q000187:drug effects; D002273:Carcinogens; D019826:Cholic Acid; D002793:Cholic Acids / Q000378:metabolism; D004058:Diffusion; D007700:Kinetics; D008074:Lipoproteins / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008563:Membrane Lipids / Q000378:metabolism*; D010100:Oxygen / Q000494:pharmacology; D010447:Peptide Hydrolases / Q000494:pharmacology; D010539:Permeability; D010590:Phalloidine / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013816:Thermodynamics
D000348:Aflatoxins; D001647:Bile Acids and Salts; D002273:Carcinogens; D002793:Cholic Acids; D008074:Lipoproteins; D008081:Liposomes; D008563:Membrane Lipids; D010590:Phalloidine; D016604:Aflatoxin B1; D010447:Peptide Hydrolases; D019826:Cholic Acid; D010100:Oxygen