Abstract
Hepatocellular transport of some substances with predominantly biliary elimination was studied by use of isolated rat liver cells. Active uptake of the hydrophilic compounds ouabain and cholate proceeded over 10–20 min and the final intracellular concentrations exceeded the extracellular concentrations more than 60-fold. Uptake of both substances was saturable and compatible with carrier-mediation. Uptake of the lipophilic compounds digitoxin and naphthol was not active, terminated within 1 min and compatible with simple diffusion.
The velocity of uptake depended on 3 major parameters: substrate features, cellular factors, and experimental conditions. Substrate features included: lipophilicity or hydrophilicity of the compound. Cellular factors included: presence and activity of transport systems. Experimental conditions included: substrate concentration, amount of cells, composition of medium.
Intracellular conjugation of cholate and naphthol produced the same polar metabolites which are formed in vivo. These metabolites were then in part released from the isolated hepatocytes into the medium. The transport systems for release are more difficult to characterize than those for uptake, and therefore little is known about their nature, despite their importance in detoxication.
If isolated hepatocytes are used in toxicology to assess metabolism or biological effects of foreign compounds, transport across the liver cell membrane is potentially a rate limiting step.
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Anwer, M. S., Hegner, D.: Effect of Na+ on bile acid uptake by isolated rat hepatocytes. Hoppe Seylers Z. Physiol. Chem. 359, 181–192 (1978)
Anwer, M. S., Hegner, D.: Study of cholic acid conjugation by isolated rat hepatocytes. Hoppe Seylers Z. Physiol. Chem. 360, 515–522 (1979)
Anwer, M. S., Kroker, R., Hegner, D.: Bile acids secretion and synthesis by isolated rat hepatocytes. Biochem. Biophys. Res. Comm. 64, 603–609 (1975)
Anwer, M. S., Kroker, R., Hegner, D.: Cholic acid uptake into isolated rat hepatocytes. Hoppe Seylers Z. Physiol. Chem. 357, 1477–1486 (1976a)
Anwer, M. S., Kroker, R., Hegner, D.: Effect of albumin on bile acid uptake by isolated rat hepatocytes. Is there a common bile acid carrier? Biochem. Biophys. Res. Commun. 73, 63–71 (1976b)
Ayala, E., Canonico, P. G.: Aminoisobutyric acid transport in primary cultures of normal adult rat hepatocytes. Proc. Soc. Exp. Biol. Med. 149, 1019–1022 (1975)
Banerjee, D., Redman, C. M.: Effect of local anaesthetics on plasma protein secretion by rat hepatocytes. Biochim. Biophys. Acta 500, 49–60 (1977)
Baur, H., Heldt, H. W.: Transport of hexoses across the liver cell membrane. Eur. J. Biochem. 74, 397–403 (1977)
Baur, H., Kasperek, S., Pfaff, E.: Criteria of viability of isolated liver cells: Hoppe Seylers Z. Physiol. Chem. 356, 827–838 (1975)
Berg, T., Iversen, J. G.: K+-transport in isolated rat liver cells stimulated by glucagon and insulin in vitro. Acta Physiol. Scand. 97, 202–208 (1976)
Berry, M. N., Friend, D. S.: High yield preparation of isolated rat liver parenchymal cells. J. Cell. Biol. 43, 506–520 (1969)
Bezooijen v., C. F., Grell, T., Knook, D. L.: Bromosulfophthalein uptake by isolated liver parenchymal cells. Biochem. Biophys. Res. Commun. 69, 354–361 (1976)
Cabantchik, Z. I., Wolosin, J. M., Ginsburg, H., Zemel, O.: Structural and functional properties of the anion transport system isolated from human erythrocyte membranes. In: Biochemistry of membrane transport. Semenza, G., Garafoli, E. (eds), pp. 328–345. Springer Berlin, Heidelberg, New York: 1977
Chen, C. P., Lee, R. H.: Active transport of alpha-aminobutyric acid in freshly prepared rat hepatocytes. Life Sci. 21, 577–583 (1977)
Drochmans, P., Wanson, J. C., May, C., Bernaert, D.: Ultrastructural and metabolic studies of isolated and cultured hepatocytes. Ciba Found. Sympos. 55, 7–24 (1977)
Dubinsky, W. P., Cockrell, R. S.: Ca++-transport across plasma and mitochondrial membranes of isolated hepatocytes. Febs Lett. 59, 39–43 (1975)
Eaton, D. L., Klaassen, C. D.: Carrier-mediated transport of ouabain in isolated hepatocytes. J. Pharmacol. Esp. Ther. 205, 480–488 (1978)
Floren, C., Nilsson, A.: Uptake and degradation of iodine-labelled chylomicron remnant particles by monolayers of rat hepatocytes. Biochem. J. 174, 827–838 (1978)
Frimmer, M.: Phalloidin, a membrane specific toxin: In: Pathogenesis and mechanisms of liver cell necrosis. Keppler, D. (ed.), pp. 163–173. Lancaster: MTP press 1975
Gardner, B., Chenouda, M. S.: Studies of bile acid secretion by isolated rat hepatocytes. J. Lipid Res. 19, 985–991 (1978)
Hardison, W. G., Weiner, R. G.: Characteristics of taurine transport in primary cultures of rat hepatocytes. Gastroenterology 76, 1283 (1979)
Home, D. W., Briggs, W. T., Wagner, C.: A functional active transport system for methotrexate in freshly isolated hepatocytes. Biochem. Biophys. Res. Commun. 68, 70–78 (1976)
Iga, T., Eaton, D. L., Klaassen, C. D.: Uptake of unconjugated bilirubin by isolated rat hepatocytes. Am. J. Physiol. 236, C9–14 (1979)
Iwamoto, K., Eaton, D. L., Klaassen, C. D.: Uptake of morphine and nalorphine by isolated rat hepatocytes. J. Pharmacol. Exp. Ther. 206, 181–189 (1978)
Jones, C. A., Moore, B. P., Cohen, G. M., Fry, J. R., Bridges, J. W.: Studies on the metabolism and excretion of benzopyrene in isolated adult rat hepatocytes. Biochem. Pharmacol. 27, 693–702 (1978)
Kelley, D. S., Potter, V. R.: Regulation of amino acid transport systems by amino acid depletion and supplementation in monolayer cultures of rat hepatocytes. J. Biol. Chem. 253, 9009–9017 (1978)
Kimmich, G. A.: Preparation and characterization of isolated intestinal epithelial cells and their use in studying intestinal transport. In: Methods in membrane biology, Vol. 5. Korn, E. D. (ed.), pp 51–115. New York: Plenum Press 1975
Klaassen, C. D.: Immaturity of the newborn rat's hepatic excretory function for ouabain. J. Pharmacol. Exp. Ther. 183, 520–526 (1972)
Kletzien, R. F., Pariza, M. W., Becker, J. E., Potter, V. R., Butcher, F. R.: Induction of amino acid transport in primary cultures of adult rat liver parenchymal cells by insulin. J. Biol. Chem. 251, 3014–3020 (1976)
Klingenberg, M., Pfaff, E.: Means of terminating reactions. Methods Enzymol. 10, 680–684 (1967)
Krenning, E. P., Docter, R., Bernard, H. F., Visser, T. J., Hennemann, G.: Active transport of trijodothyronine into isolated rat liver cells. Febs Lett. 91, 113–116 (1978)
LeCam, A., Freychet, P.: Neutral amino acid transport; characterization of the A and L systems in isolated rat hepatocytes. J. Bio. Chem. 252, 148–156 (1977)
Minder, E., Paumgartner, G.: Disparate Na+-requirement of bile acid and dye uptake by isolated hepatocytes. Arch. Hellenic Med. Soc. Suppl. 3, 12 (1977)
Nakai, T., Otto, P. S., Kennedy, D. L., Whayne, T. F.: Rat high density lipoprotein subfraction uptake and catabolism by isolated rat liver parenchymal cells. J. Biol. Chem. 251, 4914–4921 (1976)
Nilsson, Å., Akesson, B.: Uptake of chyle cholesterol esters and intact triglycerides by suspended hepatocytes. Febs Lett. 51, 219–223 (1975)
Norling, A., Moldeus, P., Andersson, B., Hänninen, O.: Passage of glucuronides and sulfates into isolated hepatocytes and action on conjugation reactions. In: Conjugation reactions in drug biotransformation. Aitio, A. (ed.), pp. 303–313. Amsterdam: Elsevier Press 1978
Olinger, E. J., Hercker, E. S., Ostrow, J. D.: Role of microtubules in bile acid transport in isolated rat hepatocytes. Gastroenterology 74, 1073 (1978)
Petzinger, E.: Comparative studies on the uptake of 14-C bile acids and 3-H demethylphalloin in isolated rat liver cells. Naunyn Schmiedeberg's Arch. Pharmacol. 307 (Suppl.), R 26, (1979)
Pfaff, E., Schuler, B., Krell, H., Höke, H.: Quality control and special features of isolated hepatocytes. Arch. Toxicol. 44, 3–21 (1980)
Potter, V. R.: Hormonal induction of enzyme functions, cyclic AMP levels and AIB transport in Morris hepatomas and in normal liver systems. In: Adv. in Expt. Med. and Biol. 92. Morris and Criss (eds.), pp. 59–88. New York: Plenum Press 1978
Rao, M. L., Rao, G. S., Höller, M., Breuer, H., Schattenberg, P. J., Stein, W.: Uptake of cortisol by isolated rat liver cells. Hoppe Seylers Z. Physiol. Chem. 357, 573–584 (1976)
Rao, M. L., Rao, G. S., Breuer, H.: Uptake of estrone, estradiol-17β and testosterone by isolated rat liver cells. Biochem. Biophys. Res. Commun. 77, 566–573 (1977)
Remmer, H.: The role of the liver in drug metabolism. Am. J. Med. 49, 617–629 (1970)
Reichen, J., Berman, M. D.: The role of microfilaments and microtubules in bile salt uptake by isolated rat hepatocytes. Gastroenterology 74, 1167 (1978)
Schwarz, L. R., Burr, R., Schwenk, M., Pfaff, E., Greim, H.: Uptake of taurocholic acid into isolated rat liver cells. Eur. J. Biochem. 55, 617–623 (1975)
Schwarz, L. R., Schwenk, M., Pfaff, E., Greim, H.: Excretion of taurocholate from isolated hepatocytes. Eur. J. Biochem. 71, 369–373 (1976)
Schwarz, L. R., Summer, K. H., Schwenk, M.: Transport and metabolism of bromosulfophtalein by isolated rat liver cells. Eur. J. Biochem. 94, 617–622 (1979a)
Schwarz, L. R., Götz, R., Klaassen, C. D.: Uptake of bromosulfophthalein-SH conjugate by isolated hepatocytes. Naunyn Schmiedeberg's Arch Pharmacol. 307 (Suppl.), R 11 (1979b)
Schwarz, L. R., Schwenk, M., Barth, C., Greim, H.: Studies with isolated liver cells: Effect of taurolithocholate on the transport of taurocholate and bromosulfophthalein. In: Biological effects of bile acids. Paumgartner G., Stiel, A. (eds.), pp. 127–133. Lancaster: MTP Press, 1979c
Schwenk, M., Wiedmann, T.: Uptake and release of ouabain by isolated rat liver cells. Hoppe Seylers Z. Physiol. Chem. 360, 373 (1979)
Schwenk, M., Burr, R., Schwarz, L. R., Pfaff, E.: Uptake of bromosulfophthalein by isolated liver cells. Eur. J. Biochem. 64, 189–197 (1976a)
Schwenk, M., Burr, R., Pfaff, E.: Influence of viability on bromosulfophthalein uptake by isolated liver cells. Naunyn Schmiedeberg's Arch. Pharmacol. 295, 99–102 (1976b)
Schwenk, M., López del Pino V., Remmer, H.: An experimental model for the study of hepatic transport and metabolism of toxic compounds. Arch. Toxicol. Suppl. 2, 339–343 (1979a)
Schwenk, M., López del Pino V., Bolt, H. M.: The kinetics of hepatocellular transport and metabolism of estrogens. J. Steroid Biochem. 10, 37–41 (1979b)
Ungemach, F. R., Hegner, D.: Uptake of thymidine into isolated rat hepatocytes. Hoppe Seylers Z. Physiol. Chem. 359, 845–856 (1978)
Usui, T.: Thin layer chromatography of bile acids with special reference to separation of keto bile acids. J. Biochem. (Tokyo) 54, 283–286 (1963)
Vonk, R. J., Jekel, P. A., Meier, D. K. F., Hardonk, M. J.: Transport of drugs in isolated hepatocytes, the influence of bile salts. Biochem. Pharmacol. 27, 397–405 (1978)
Wiedmann, T.: Membrane transport of cardiac glycosides by isolated rat hepatocytes. Naunyn Schmiedeberg's Arch. Pharmacol. 307, (Suppl.) R 12 (1979)
Wisher, M. H., Evans, W. H.: Enzymes of the hepatocyte plasma membrane. In: Membrane alterations as basis of liver injury. Popper, H. (ed.), pp. 127–141. Lancaster: MTP press 1977
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Schwenk, M. Transport systems of isolated hepatocytes. Arch. Toxicol. 44, 113–126 (1980). https://doi.org/10.1007/BF00303188
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DOI: https://doi.org/10.1007/BF00303188