Abstract
Purpose. To investigate the direct effects of sex hormones, growth hormone, thyroid hormones and dexamethasone on the regulation of UDP-glucuronosyltransferase (UGT).
Methods. Rat hepatocytes were cultured on matrigel and treated with various hormones. Northern blot analysis was carried out using cDNA probes to family 1 and family 2 isoforms.
Results. Treatment with 10−5 M testosterone increased the mRNA levels of UGT 2B1 by 29% and UGT2B3 by 32%. Incubation of growth hormone (10 mU) with hepatocytes suppressed the expression of UGT2B1 and UGT2B3 by 17% and 38%, respectively. T3 administration resulted in a time and dose-dependent effect on the expression of UGT 1 isoforms, with increased UGT1A6 by 70%, and decreased UGT1A1 by 38% and UGT1A5 by 35%. All UGT isoforms except UGT 1A6 studied in this assay were up-regulated by dexamethasone, but to different degrees. The regulation of UGT1A1 and UGT2B1 by dexamethasone was dose and time dependent, and the induction of dexamethasone in the expression of UGT1A1 and UGT2B1 was blocked by cycloheximide but not dichloro-1-D-ribofuranosylbenzimidazole.
Conclusions. This study demonstrates that multiple hormones take part in the regulation of UGT mRNA expression in the rat and individual genes can be differentially modulated.
Similar content being viewed by others
REFERENCES
P. I. Mackenzie, I. S. Owens, B. Burchell, K. W. Bock, A. Bairoch, A. Belanger, S. Fournel-Gigleux, M. Green, D. W. Hum, T. Iyanagi, D. Lancet, P. Louisot, J. Magdalou, J. R. Chowdhury, J. K. Ritter, H. Schachter, T. R. Tephly, K. F. Tipton, and D. W. Nebert. The UDP glycosyltransferase gene superfamily: recommended nomenclature update based on evolutionary divergence. Pharmacogenetics 7:255–269 (1997).
G. S. Rao, G. Haueter, M. L. Rao, and H. Breuer. Steroid glucuronyltransferases of rat liver. Properties of oestrone and testosterone glucuronyltransferases and the effect of ovariectomy, castration and administration of steroids on the enzymes. Biochem. J. 162:545–556 (1977).
M. Muraca and J. Fevery. Influence of sex and sex steroids on bilirubin uridine diphosphate-glucuronosyltransferase activity of rat liver. Gastroenterology 87:308–313 (1984).
S. I. Strasser, S. A. Smid, M. L. Mashford, and P. V. Desmond. Sex hormones differentially regulate isoforms of UDP-glucuronosyltransferase. Pharm. Res. 14:1115–1121 (1997).
T. Masmoudi, R. Planells, J. Mounie, Y. Artur, J. Magdalou, and H. Goudonnet. Opposite regulation of bilirubin and 4-nitrophenol UDP-glucuronosyltransferase mRNA levels by 3,3′,5 triiodo-L-thyronine in rat liver. FEBS Lett 379:181–185 (1996).
Y. Emi, S. Ikushiro, and T. Iyanagi. Drug-responsive and tissue-specific alternative expression of multiple first exons in rat UDP-glucuronosyltransferase family 1 (UGT1) gene complex. J. Biochem. (Tokyo) 117:392–399 (1995).
D. M. Bissell and P. S. Guzelian. Phenotypic stability of adult rat hepatocytes in primary monolayer culture. Ann. N Y Acad. Sci. 349:85–98 (1980).
C. Liddle, A. Mode, C. Legraverend, and J. A. Gustafsson. Constitutive expression and hormonal regulation of male sexually differentiated cytochromes P450 in primary cultured rat hepatocytes. Arch. Biochem. Biophys. 298:159–166 (1992).
P. Chomczynski and N. Sacchi. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162:156–159 (1987).
P. I. Mackenzie. Rat liver UDP-glucuronosyltransferase. Sequence and expression of a cDNA encoding a phenobarbital-inducible form. J. Biol. Chem. 261:6119–6125 (1986).
P. I. Mackenzie. Rat liver UDP-glucuronosyltransferase. Identification of cDNAs encoding two enzymes which glucuronidate testosterone, dihydrotestosterone, and beta-estradiol. J. Biol. Chem. 262:9744–9749 (1987).
Y. Emi, S. Ikushiro, and T. Iyanagi. Xenobiotic responsive element-mediated transcriptional activation in the UDP-glucuronosyltransferase family 1 gene complex. J. Biol. Chem. 271:3952–3958 (1996).
O. Weijtens, R. C. Schoemaker, A. F. Cohen, F. P. Romijn, E. G. Lentjes, J. van Rooij, and J. C. van Meurs. Dexamethasone concentration in vitreous and serum after oral administration. Am. J. Ophthalmol. 125:673–679 (1998).
K. Nawa, T. Nakamura, A. Kumatori, C. Noda, and A. Ichihara. Glucocorticoid-dependent expression of the albumin gene in adult rat hepatocytes. J. Biol. Chem. 261:16883–16888 (1986).
T. Nakamura, S. Niimi, K. Nawa, C. Noda, A. Ichihara, Y. Takagi, M. Anai, and Y. Sakaki. Multihormonal regulation of transcription of the tryptophan 2,3-dioxygenase gene in primary cultures of adult rat hepatocytes with special reference to the presence of a transcriptional protein mediating the action of glucocorticoids. J. Biol. Chem. 262:727–733 (1987).
C. J. Pan, L. L. Shelly, D. S. Rabin, and J. Y. Chou. Inhibition of tyrosine aminotransferase gene expression by retinoic acid. Mol. Endocrinol. 6:572–580 (1992).
L. J. Hager and R. D. Palmiter. Transcriptional regulation of mouse liver metallothionein-I gene by glucocorticoids. Nature 291:340–342 (1981).
S. R. Spindler, S. H. Mellon, and J. D. Baxter. Growth hormone gene transcription is regulated by thyroid and glucocorticoid hormones in cultured rat pituitary tumor cells. J. Biol. Chem. 257:11627–11632 (1982).
F. Payvar, D. DeFranco, G. L. Firestone, B. Edgar, O. Wrange, S. Okret, J. A. Gustafsson, and K. R. Yamamoto. Sequence-specific binding of glucocorticoid receptor to MTV DNA at sites within and upstream of the transcribed region. Cell 35:381–392 (1983).
R. M. Evans and S. M. Hollenberg. Cooperative and positional independent transactivation domains of the human glucocorticoid receptor. Cold Spring Harb. Symp. Quant. Biol. 53 Pt 2:813–818 (1988).
V. A. Catania, A. J. Dannenberg, M. G. Luquita, E. J. Sanchez Pozzi, J. K. Tucker, E. K. Yang, and A. D. Mottino. Gender-related differences in the amount and functional state of rat liver UDP-glucuronosyltransferase. Biochem. Pharmacol. 50:509–514 (1995).
N. Nemoto and J. Sakurai. Glucocorticoid and sex hormones as activating or modulating factors for expression of Cyp2b-9 and Cyp2b-10 in the mouse liver and hepatocytes. Arch. Biochem. Biophys. 319:286–292 (1995).
P. S. Guzelian, D. Li, E. G. Schuetz, P. Thomas, W. Levin, A. Mode, and J. A. Gustafsson. Sex change in cytochrome P-450 phenotype by growth hormone treatment of adult rat hepatocytes maintained in a culture system on matrigel. Proc. Natl. Acad. Sci. USA 85:9783–9787 (1988).
F. Gueraud, T. Masmoudi, H. Goudonnet, and A. Paris. Differential effect of hypophysectomy and growth hormone treatment on hepatic glucuronosyltransferases in male rats: evidence for an action at a pretranslational level for isoforms glucuronidating bilirubin. Biochem. Pharmacol. 53:1637–1647 (1997).
M. Ciotti, M. T. Yeatman, R. J. Sokol, and I. S. Owens. Altered coding for a strictly conserved di-glycine in the major bilirubin UDP-glucuronosyltransferase of a Crigler-Najjar type I patient. J. Biol. Chem. 270:3284–3291 (1995).
M. K. Bhat, C. I. Yu, N. Yap, Q. Zhan, Y. Hayashi, P. Seth, and S. Cheng. Tumor suppressor p53 is a negative regulator in thyroid hormone receptor signaling pathways. J. Biol. Chem. 272:28989–28993 (1997).
T. Masmoudi, A. K. Hihi, M. Vazquez, Y. Artur, B. Desvergne, W. Wahli, and H. Goudonnet. Transcriptional regulation by triiodothyronine of the UDP-glucuronosyltransferase family 1 gene complex in rat liver. Comparison with induction by 3-methylcholanthrene. J. Biol. Chem. 272:17171–17175 (1997).
H. Goudonnet, J. Magdalou, J. Mounie, A. Naoumi, M. L. Viriot, A. Escousse, G. Siest, and R. Truchot. Differential action of thyroid hormones and chemically related compounds on the activity of UDP-glucuronosyltransferases and cytochrome P-450 isozymes in rat liver. Biochim. Biophys. Acta 1035:12–19 (1990).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Li, V.Q., Prentice, D.A., Howard, M.L. et al. The Effect of Hormones on the Expression of Five Isoforms of UDP-Glucuronosyltransferase in Primary Cultures of Rat Hepatocytes. Pharm Res 16, 191–197 (1999). https://doi.org/10.1023/A:1018812021549
Issue Date:
DOI: https://doi.org/10.1023/A:1018812021549