Role of Human Hepatocyte Nuclear Factor 4α in the Expression of Drug-Metabolizing Enzymes and Transporters in Human Hepatocytes Assessed by Use of Small Interfering RNA

doi:10.2133/dmpk.22.287

Drug Metabolism and Pharmacokinetics

Volume 22, Issue 4, 2007, Pages 287-298

https://doi.org/10.2133/dmpk.22.287 Get rights and content

Summary:

Hepatocyte nuclear factor 4α (HNF4α) is an important transcription factor in hepatic gene expression. Here, we have investigated the role of HNF4α in the expression of drug-metabolizing enzymes and transporters in human hepatocytes using an adenovirus expressing human HNF4α-small interfering RNA (hHNF4α-siRNA). The hHNF4α-siRNA effectively reduced the mRNA and nuclear protein levels of hHNF4α in a concentration-dependent manner. The hHNF4α-siRNA also decreased the mRNA levels of CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A4, UGT1A1, UGT1A9, SULT2A1, ABCB1, ABCB11, ABCC2, OATP1B1 and OCT1, as well as those of PXR and CAR. To discern the role of these nuclear receptors, we co-infected hepatocytes with hHNF4α-siRNA and PXR- or CAR-expressing adenovirus. The hHNF4α-siRNA-induced reductions of the enzyme and transporter mRNA levels were not restored except CYP2B6 mRNA levels, which were returned to the control level by overexpressing CAR. Furthermore, although hHNF4α-siRNA did not significantly affect the fold-induction of CYP2B6, CYP2C8, CYP2C9, or CYP3A4 mRNA levels following treatment with CYP inducers, the levels in hHNF4α-suppressed cells fell significantly compared to the control. These results suggest that HNF4α plays a dominant role in the expression of drug-metabolizing enzymes and transporters in human hepatocytes, and that HNF4α expression levels is a possible determinant for inter-individual variations in the expression of these enzymes and transporters.

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Citation Excerpt :
This not only lowers the cost but more importantly reduces the potential toxicity issues derived from said combination. HNF40 is known to regulate the expression of a number of drug metabolizing enzymes and transporters [47,48]. Glycyrrhizin may exert its “drug assistant” effect by suppressing the expression of these drug metabolizing enzymes and transporters under the control of HNF4α; thus, reducing the dosage necessary and modulating the half-life of active ingredients derived from said herbal medicine to achieve sufficient therapeutic exposure.
Hepatocyte nuclear factor 4 alpha (HNF4α) regulates the expression of essential genes involved in very-low-density lipoprotein (VLDL) homeostasis and gluconeogenesis.　18β-glycyrrhetinic acid (GA) is an active ingredient of Glycyrrhiza uralensis an herbal medicine used for treating liver aliments. In this study, we established that GA functions as a partial antagonist of HNF4α through HNF4α-driven reporter luciferase assay and co-immunoprecipitation experiments with co-activator PGC1α. By virtual docking and site-directed mutagenesis analysis, we confirmed that serine 190 and arginine 235 of HNF4α are both essential for GA to exert its antagonistic action on HNF4α. Importantly, GA suppressed the expression of HNF4α target genes such as apolipoprotein B (ApoB), microsomal triglyceride transfer protein (MTP) and phospholipase A₂ G12B (PLA2G12B) modulating hepatic VLDL secretion in mice fed on a high fat diet. In addition, GA also suppressed gluconeogenesis and ameliorated glucose intolerance via down-regulating the expression of HNF4α target genes glucose-6-phosphatase (G6pc) and phosphoenolpyruvate carboxykinase (Pepck). Furthermore, GA significantly lowered blood glucose and improved insulin resistance in db/db mice. In all, we established that GA acts as a partial HNF4α antagonist modulating lipid and carbohydrate metabolism.

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Regular ArticleRole of Human Hepatocyte Nuclear Factor 4α in the Expression of Drug-Metabolizing Enzymes and Transporters in Human Hepatocytes Assessed by Use of Small Interfering RNA

Summary:

J. Biol. Chem.

Hepatology

Arch. Biochem. Biophys.

Drug Metab. Pharmacokinet.

Drug Metab. Pharmacokinet.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

FEBS Lett.

A potential role for antagonistic interactions between members of the nuclear receptor family. J. Biol. Chem.

Hepatology

J. Biol. Chem.

J. Biol. Chem.

Liver-enriched transcription factor HNF-4 is a novel member of the steroid hormone receptor superfamily

Genes Dev.

Orphan receptor HNF-4 and liver-specific gene expression

Receptor

Hepatocyte nuclear factor 4 expression overcomes repression of the hepatic phenotype in dedifferentiated hepatoma cells

Mol. Cell Biol.

Liver-enriched transcription factors and hepatocyte differentiation

FASEB J.

The maturity-onset diabetes of the young (MODY1) transcription factor HNF4α regulates expression of genes required for glucose transport and metabolism

Proc. Natl. Acad. Sci. USA

Hepatocyte nuclear factor 4α (nuclear receptor 2A1) is essential for maintenance of hepatic gene expression and lipid homeostasis

Mol. Cell Biol.

Mutations in the hepatocyte nuclear factor-4α gene in maturity-onset diabetes of the young (MODY1)

Nature

Regular Article
Role of Human Hepatocyte Nuclear Factor 4α in the Expression of Drug-Metabolizing Enzymes and Transporters in Human Hepatocytes Assessed by Use of Small Interfering RNA