In this study, ethinylestradiol inhibited the uptake of taurocholate by cultured rat hepatocytes, increasing the Km1 while leaving the Vmax unchanged. S-AdenosylL-methionine (SAMe) had no effect on taurocholate uptake or release, but was able to reverse the competitive inhibition induced by ethinylestradiol. S-Adenosyl-L-homocysteine did not reverse this inhibition, which suggests that the methyl group of SAMe affects its activity. Several possible mechanisms for the action of SAMe were investigated. The methylation of cell membrane phospholipids was eliminated as a possible mechanism. The presence of SAMe greatly increased the catabolism of ethinylestradiol by hepatocytes and reduced its covalent binding to hepatocyte macromolecules. In culture supernatants, both highly polar (conjugated) and non-conjugated metabolites could be detected. Moreover, most of the metabolites were methylated. This suggests that SAMe may revert the effects of ethinylestradiol of taurocholate uptake by increasing its catabolic rate by hepatocytes.
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Abbreviations
- SAMe,:
-
S-adenosyl-L-methionine
- EE,:
-
17α-ethinylestradiol
References
FADACHI, Y., NANNO, T., and KANBE., A. (1986). “The effects of S-adenosyl methionine on intrahepatic cholestasis.” Jpn. Arch. Int. Med. 6:185–192.
ARIAS, A.M., ADACHI, Y., and TRAN, T. (1983). “Ethinyl estradiol cholestasis: a disease of the sinusoidal domain of hepatocyte plasma membrane.” Hepatology 3:872.
BERR, F., SIMON, F.R., and REICHEN, J. (1987). “Ethinylestradiol impairs bile salt uptake and Na-K pump function of rat hepatocytes.” Am. J. Physiol. 247:G437.
BOELSTERLI, U.A., RAKHIT, G., and BALAZS, T. (1983). “Modulation by S-adenosyl-Lmethionine of hepatic Na+/K−-ATPase, membrane fluidity, and bile flow in rats with ethinyl estradiol-induced cholestasis.” Hepatology 3:12–17.
BOYER, J.L. (1980). “New concepts of mechanisms of hepatocyte bile formation.” Physiol. Rev. 60:302–326.
BOYER, J.L. (1980). “New concepts of mechanisms of hepatocyte bile formation.” Physiol Rev 60:302–326.
BOYER, J.L., ALLEN, R.M., and NG, O.C. (1983). “Biochemical separation of Na+, K+-ATPase from a purified light density, canalicular enriched plasma membrane fraction from rat liver.” Hepatology 3:18–28.
BROCK, W.J., DURHAM, S., and VORE, M. (1984). “Characterization of the interaction between estrogen metabolites and taurocholate for uptake into isolated hepatocytes. Lack of correlation between cholestasis and inhibition of taurocholate uptake.” J. Steroid Biochem. 20:1181–1185.
BROWN, D., DUDEJA, P.K., and BRASITUS, T.A. (1988). “S-Adenosyl-L-methionine modulates Na+/K+ ATPase activity in rat colonic basolateral membranes.” Biochem. J 251:215–222.
CHIARANTI, E., ARCANGELI, A., and MAZZANTI, R. (1979). “Ethinyl estradiol induced cholestasis. Morphological and functional aspects.” In: Problems in intrahepatic cholestasis. Second International Symposium. Florence, Italy, 1978. Karger, Basel, Switzerland. pp. 102–110.
DAVID, G., OELBERG, D.G., and LESTER, R. (1986). “Cellular mechanisms of cholestasis.” Ann. Rev. Med. 37:297–317.
DAVIS, R.A., KERN, F., SHOWALTER, R., SUTHERLAND, E., SINESKY, M., and SIMON, F.R. (1978). “Alterations of hepatic Na+, K+-ATPase and bile flow by estrogen: Effects on liver surface membrane lipid structure and function.” Proc. Natl. Acad. Sci. 75:4130–4134.
DI PADOVA, C., TRITAPE, R., DI PADOVA, F., FREZZA, M., and STRAMENTINOLI, G. (1984). “S-Adenosyl-L-Methionine antagonizes oral contraceptive-induced bile cholesterol supersaturation in healthy women: preliminary report of controlled randomized trial.” Am. J. Gastroenterol. 79:941–944.
DONATO, M.T., CASTELL, J.V., and GóMEZ-LECHóN, M.J. (1991). “Co-cultures of hepatocytes with epithelial-like cell lines: Expression of drug-biotransformation activities by hepatocytes.” Cell Biol. Toxicol. 7:1–14.
ELIAS, E. and BOYER, J.L. (1979). “Mechanisms of intrahepatic cholestasis.” In: Progress in liver diseases. Vol. 6. (H. Popper and F. Schaffner, eds.). Grune & Stratton, New York, NY. pp. 457–470.
FREZZA, M., CAMMARERI, G., and DI PADOVA, C. (1987a). “Beneficial effect of S-Adenosyl-LMethionine in pregnant women with cholestasis: results of a multicenter controlled clinical trial.” J. Hepatol. 5:S27.
FREZZA, M., DI PADOVA, C., and THE ITALIAN STUDY GROUP FOR SAME IN LIVER DISEASE. (1987b). “Multicenter placebo controlled trial of intravenous and oral S-Adenosylmethionine (SAMe) in cholestatic patients with liver disease.” Hepatology 7:1105.
GóMEZ-LECHóN, M.J. and CASTELL, J.V. (1983). “The role of fetal calf serum during the first stages of hepatocyte culture.” In: Hormonally defined media: A tool in cell biology (G. Fisher and R.J. Wieseer, eds.). Springer-Verlag, Berlin, Germany. pp. 340–343.
GóMEZ-LECHóN, M.J., LOPEZ, P., and CASTELL, J.V. (1984). “Biochemical functionality and recovery of hepatocytes after deep-freezing storage.” In Vitro 20:826–832.
ISHAK, K.G. and ZIMMERMAN, H.J. (1987). “Hepatotoxic effects of the anabolic/androgenic steroids.” Sem. Liv. Dis. 7:230–236.
KIMELBERG, H.K. (1975). “Alterations in phospholipid-dependent Na+, K+-ATPase activity due to lipid fluidity. Effects of cholesterol and Mg+.” Biochem. Biophys. Acta. 413:143–156.
KLAASEN, C.D. and WATKINS, J.B. (1984). “Mechanisms of bile formation, hepatic uptake, and biliary excretion.” Pharamacol. Rev. 36:1–67.
KULKARINI, B.D. and GOLDIZIEHER, J.W. (1970). “Preliminary information about urinary excretion samples and the isolation method of 14C-ethinylestradiol metabolites in women.” Contraception 1:1–47.
LOWRY, O.H., ROSENBROUGH, N.J., FARR, A.L., and RANDALL, R.J. (1951). “Protein measurement with the Folin phenol reagent.” J. Biol. Chem. 193:265–275.
PELECH, S.L. and VANCE, D.E. (1984). “Regulation of phosphatidylcholine biosynthesis.” Biochim. Biophys. Acta. 779:217–251.
REICHEN, J. and SIMON, F.R. (1982). “Cholestasis.” In: The liver: Biology and pathobiology (I. Arias, H. Popper, D. Schachter, and D.A. Shafritz, eds.). Raven Press, New York, NY. pp. 785–800.
SCHREIBER, A.J. and SIMON, F.R. (1983). “Estrogen-induced cholestasis: Clues to pathogenesis and treatment.” Hepatology 4:607–71.
SIMON, F.R., GONZALEZ, M., SUTHERLAND, E., ACCATINO, L., and DAVIS, R.A. (1987). “Reversal of ethinyl estradiol-induced bile secretory failure with triton WR-1339.” J. Clin. Invest. 65:851–860.
STACEY, N.H. (1986). “Effects of ethinyl estradiol on substrate uptake and efflux by isolated rat hepatocytes.” Biochem Pharmacol 35:2495–2500.
STRAMENTINOLI, G., GUALANO, M., and DI PADOVA, C. (1977). “Effect of S-adenosyl-Lmethionine on ethinylestradiol-induced impairment of bile flow in female rats.” Experientia 33:1361–1362.
STRAMENTINOLI, G., GUALANO, M., ROGAVNATI, P., and DI PADOVA, C. (1979). “Influence of S-Adenosyl-L-Methionine on irreversible binding of ethinyl estradiol to rat liver microsomes, and its implication in bile secretion.” Biochem. Pharmacol. 28:981–984.
STRAMENTINOLI, G., DI PADOVA, C., GUALANO, tM., ROGAVNATI, P., and GALLIKIENLE, M. (1981). “Ethinyl estradiol-induced impairment of bile secretion in the rat: protective effects of S-adenosyl-L-methionine and its implication in estrogen metabolism.” Gastroenterology 80:154–158.
TALAAT ABDEL-AZIZ, M. and WILLIAMS, K. (1970). “The metabolism of the radioactive 17α-ethinylestradiol in woman.” Steroids 15:695–710.
TOWELL, D.A. (1942). “Isolated liver perfusion in the study of hepatic function.” J. Physiol. 100:432–435.
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Larrauri, A., Castell, J.V., Garrido, G. et al. S-adenosyl-L-methionine reverses the cholestatic effect of ethinylestradiol in rat hepatocytes by increasing its catabolism. Cell Biol Toxicol 8, 13–26 (1992). https://doi.org/10.1007/BF00119292
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DOI: https://doi.org/10.1007/BF00119292