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PMID: 16799975 Published · ppublish English Comparative Study Journal Article Research Support, N.I.H., Extramural

Phenobarbital regulates nuclear expression of HNF-4alpha in mouse and rat hepatocytes independent of CAR and PXR.

Hepatology (Baltimore, Md.) ·Vol. 44 ·No. 1 ·2006-07-00 ·Pages 186-94

Bell AW, Michalopoulos GK

Abstract

Phenobarbital is a lipophilic molecule used as a sedative and antiepileptic drug that elicits a multitude of effects in the liver, including gross liver enlargement, hepatocyte hypertrophy, and induced expression of drug-metabolizing enzymes and other liver-specific genes. The constitutive androstane receptor (CAR; NR1I3) and to a lesser extent the pregnane X receptor (PXR; NR1I2) are responsible for mediating induction of many phenobarbital-responsive genes. However, CAR-mediated transcriptional control of some genes is critically dependent on hepatocyte nuclear factor 4 alpha (HNF-4alpha; NR2A1), which itself regulates multiple liver-specific genes involved in hepatic growth, metabolism, and differentiation. We studied the effects of phenobarbital on HNF-4alpha expression in hepatocytes and provide evidence that HNF-4alpha nuclear expression is regulated in response to phenobarbital. Real-time polymerase chain reaction analyses revealed that HNF-4alpha mRNA is modestly up-regulated by phenobarbital. In addition, nuclear expression of HNF-4alpha protein is significantly elevated 3 hours after the administration of phenobarbital in wild-type, CAR-/-, and CAR-/-/PXR-/- mice. In vitro analysis revealed that phenobarbital-induced HNF-4alpha expression is both time- and dose dependent. In addition, the phosphatase inhibitor okadaic acid and the Ca2+/calmodulin-dependent protein kinase II inhibitor KN62 block nuclear induction of HNF-4alpha by phenobarbital. Furthermore, HNF-4alpha nuclear expression is enhanced by inhibition of cyclic AMP-dependent protein kinase A. In conclusion, induced nuclear expression of HNF-4alpha and CAR is an integral part of the phenobarbital response, aimed at coordinated regulation of genes involved in drug metabolism and detoxification as well as maintenance of liver function.

MeSH Terms
Animals Anticonvulsants/pharmacology Blotting, Western Cell Nucleus/metabolism Cells, Cultured Constitutive Androstane Receptor Gene Expression/drug effects Hepatocyte Nuclear Factor 4/drug effects,genetics,metabolism Hepatocytes/cytology,drug effects,metabolism In Vitro Techniques Male Mice Mice, Inbred C57BL Phenobarbital/pharmacology Polymerase Chain Reaction Pregnane X Receptor RNA, Messenger/genetics Rats Rats, Inbred F344 Receptors, Cytoplasmic and Nuclear/metabolism Receptors, Steroid/metabolism Transcription Factors/metabolism
Chemicals
Anticonvulsants Constitutive Androstane Receptor Hepatocyte Nuclear Factor 4 Hnf4a protein, mouse Hnf4a protein, rat Nr1i2 protein, mouse Nr1i2 protein, rat Nr1i3 protein, mouse Nr1i3 protein, rat Pregnane X Receptor RNA, Messenger Receptors, Cytoplasmic and Nuclear Receptors, Steroid Transcription Factors Phenobarbital
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Bell Aaron W
Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Michalopoulos George K
References (36)
36 references, click to expand
  1. Phenobarbital-elicited activation of nuclear receptor CAR in induction of cytochrome P450 genes.
    Biochem Biophys Res Commun. 2000 Oct 14;277(1):1-6 PMID: 11027630
  2. Acetylation regulates transcription factor activity at multiple levels.
    Mol Cell. 2000 Apr;5(4):745-51 PMID: 10882110
  3. Identification of phenobarbitone-modulated genes in mouse liver by differential display.
    J Biochem Mol Toxicol. 2000;14(2):65-72 PMID: 10630419
  4. Human CYP2C8 is transcriptionally regulated by the nuclear receptors constitutive androstane receptor, pregnane X receptor, glucocorticoid receptor, and hepatic nuclear factor 4alpha.
    Mol Pharmacol. 2005 Sep;68(3):747-57 PMID: 15933212
  5. Hepatic expression of the UGT1A9 gene is governed by hepatocyte nuclear factor 4alpha.
    Mol Pharmacol. 2005 Jan;67(1):241-9 PMID: 15470081
  6. The nuclear receptors constitutive androstane receptor and pregnane X receptor cross-talk with hepatic nuclear factor 4alpha to synergistically activate the human CYP2C9 promoter.
    J Pharmacol Exp Ther. 2005 Sep;314(3):1125-33 PMID: 15919766
  7. Regulation of phenobarbital-induction of CYP2B and CYP3A genes in rat cultured hepatocytes: involvement of several serine/threonine protein kinases and phosphatases.
    Cell Biol Toxicol. 2000;16(5):325-37 PMID: 11201056
  8. Cytochrome P450 regulation by hepatocyte nuclear factor 4 in human hepatocytes: a study using adenovirus-mediated antisense targeting.
    Hepatology. 2001 Mar;33(3):668-75 PMID: 11230748
  9. Diverse roles of the nuclear orphan receptor CAR in regulating hepatic genes in response to phenobarbital.
    Mol Pharmacol. 2002 Jan;61(1):1-6 PMID: 11752199
  10. Loss of interleukin 6 results in delayed mammary gland involution: a possible role for mitogen-activated protein kinase and not signal transducer and activator of transcription 3.
    Mol Endocrinol. 2002 Dec;16(12):2902-12 PMID: 12456808
  11. The orphan nuclear receptor HNF4alpha determines PXR- and CAR-mediated xenobiotic induction of CYP3A4.
    Nat Med. 2003 Feb;9(2):220-4 PMID: 12514743
  12. Serine/threonine phosphorylation regulates HNF-4alpha-dependent redox-mediated iNOS expression in hepatocytes.
    Am J Physiol Cell Physiol. 2003 Apr;284(4):C1090-9 PMID: 12466152
  13. Role of the hepatocyte nuclear factor 4alpha in control of the pregnane X receptor during fetal liver development.
    Hepatology. 2003 Jun;37(6):1375-84 PMID: 12774017
  14. Identification of the nuclear receptor CAR:HSP90 complex in mouse liver and recruitment of protein phosphatase 2A in response to phenobarbital.
    FEBS Lett. 2003 Jul 31;548(1-3):17-20 PMID: 12885400
  15. CAR, driving into the future.
    Mol Endocrinol. 2004 Jul;18(7):1589-98 PMID: 14988430
  16. Phenobarbital-induced synthesis of the microsomal drug-metabolizing enzyme system and its relationship to the proliferation of endoplasmic membranes. A morphological and biochemical study.
    J Cell Biol. 1965 Jun;25(3):627-39 PMID: 4378769
  17. Studies on liver regeneration. II. Effects of phenobarbital on the onset and pattern of rat liver regeneration following partial hepatectomy.
    Cell Tissue Kinet. 1971 Nov;4(6):529-37 PMID: 4947050
  18. Liver growth after partial hepatectomy: influence of phenobarbital administration.
    Exp Mol Pathol. 1981 Apr;34(2):216-25 PMID: 6162677
  19. Controlled death (apoptosis) of normal and putative preneoplastic cells in rat liver following withdrawal of tumor promoters.
    Carcinogenesis. 1984 Apr;5(4):453-8 PMID: 6231134
  20. The molecular biology of cytochrome P450s.
    Pharmacol Rev. 1988 Dec;40(4):243-88 PMID: 3072575
  21. Enzyme induction in the cytochrome P-450 system.
    Pharmacol Ther. 1990;45(2):241-98 PMID: 2405437
  22. Liver tumor promotion: effect of phenobarbital on EGF and protein kinase C signal transduction and transforming growth factor-beta 1 expression.
    Dig Dis Sci. 1991 May;36(5):659-68 PMID: 2022168
  23. Effect of 2% dimethyl sulfoxide on the mitogenic properties of epidermal growth factor and hepatocyte growth factor in primary hepatocyte culture.
    J Cell Physiol. 1991 May;147(2):274-80 PMID: 1828251
  24. Cytoplasmic localization of pregnane X receptor and ligand-dependent nuclear translocation in mouse liver.
    J Biol Chem. 2004 Nov 19;279(47):49307-14 PMID: 15347657
  25. Involvement of cyclic nucleotide- and calcium-regulated pathways in phenobarbital-induced cytochrome P-450 3A expression in mouse primary hepatocytes.
    J Pharmacol Exp Ther. 1999 Sep;290(3):1270-7 PMID: 10454503
  26. Concise review of the cytochrome P450s and their roles in toxicology.
    Toxicol Sci. 1999 Apr;48(2):151-6 PMID: 10353305
  27. The quantitative distinction of hyperplasia from hypertrophy in hepatomegaly induced in the rat liver by phenobarbital.
    Toxicol Sci. 1998 Jul;44(1):46-51 PMID: 9720140
  28. The nuclear orphan receptor CAR-retinoid X receptor heterodimer activates the phenobarbital-responsive enhancer module of the CYP2B gene.
    Mol Cell Biol. 1998 Oct;18(10):5652-8 PMID: 9742082
  29. Dose-dependent biphasic effects of phenobarbital on growth and differentiation of primary culture rat hepatocytes.
    J Gastroenterol Hepatol. 1998 Sep;13 Suppl:S78-82 PMID: 9792038
  30. Phenobarbital suppresses growth and accelerates restoration of differentiation markers of primary culture rat hepatocytes in the chemically defined hepatocyte growth medium containing hepatocyte growth factor and epidermal growth factor.
    Exp Cell Res. 1998 Jun 15;241(2):445-57 PMID: 9637786
  31. Protein kinase A-dependent phosphorylation modulates DNA-binding activity of hepatocyte nuclear factor 4.
    Mol Cell Biol. 1997 Aug;17(8):4208-19 PMID: 9234678
  32. Serine/threonine phosphorylation of orphan receptor hepatocyte nuclear factor 4.
    Arch Biochem Biophys. 1997 Apr 1;340(1):1-9 PMID: 9126270
  33. Subchronic effects of dieldrin and phenobarbital on hepatic DNA synthesis in mice and rats.
    Fundam Appl Toxicol. 1996 Feb;29(2):219-28 PMID: 8742319
  34. Transforming growth factor-beta receptors type I, II and III in phenobarbital-promoted rat liver tumors.
    Carcinogenesis. 1994 Dec;15(12):2763-7 PMID: 8001232
  35. Phenobarbital selectively promotes initiated cells with reduced TGF beta receptor levels.
    Carcinogenesis. 1996 Jan;17(1):171-4 PMID: 8565130
  36. Population expansion, clonal growth, and specific differentiation patterns in primary cultures of hepatocytes induced by HGF/SF, EGF and TGF alpha in a chemically defined (HGM) medium.
    J Cell Biol. 1996 Mar;132(6):1133-49 PMID: 8601590
Article Info
Journal
Hepatology (Baltimore, Md.)
Abbr.
Hepatology
ISSN
0270-9139
DOI
10.1002/hep.21234
Published
2006-07-00
Pages
186-94
Language
English
Region
United States
NLM ID
8302946
PMCID
PMC1769339
Subset
IM
Grants
NCI NIH HHS · R01 CA035373 · United States
NCI NIH HHS · R01 CA103958 · United States
NCI NIH HHS · CA103958 · United States
NCI NIH HHS · CA35373 · United States
External Links
PubMed source DOI: 10.1002/hep.21234 PMC Full Text View journal details
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