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Bioactive Terpenoids and Flavonoids from Ginkgo Biloba Extract Induce the Expression of Hepatic Drug-Metabolizing Enzymes Through Pregnane X Receptor, Constitutive Androstane Receptor, and Aryl hydrocarbon Receptor-Mediated Pathways

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Abstract

Purpose

The objective of the current study is to investigate the hypothesis that bioactive terpenoids and flavonoids of Ginkgo biloba extract (GBE) induce human hepatic drug metabolizing enzymes (DMEs) and transporters through the selective activation of pregnane X receptor (PXR), constitutive androstane receptor (CAR), and aryl hydrocarbon receptor (AhR).

Methods

Human primary hepatocyte (HPH), and HepG2 cells are used as in vitro models for enzyme induction and nuclear receptor activation studies. A combination of real-time RT-PCR, transient transfection, and cell-based reporter assays were employed.

Results

In human primary hepatocytes, real-time PCR analysis showed induction of CYP2B6, CYP3A4, UGT1A1, MDR1, and MRP2 by EGb 761, ginkgolide A (GA) and ginkgolide B (GB), but not by bilobalide (BB) or the flavonoids (quercetin, kaempferol and tamarixetin) of GBE. Cell-based reporter assays in HepG2 revealed that GA and GB are potent activators of PXR; quercetin and kaempferol activate PXR, CAR, and AhR, whereas BB exerts no effects on these xenobiotic receptors. Notably, the flavonoids induced the expression of UGT1A1 and CYP1A2 in HepG2 cells but not in HPH.

Conclusion

Our results indicate that terpenoids and flavonoids of GBE exhibit differential induction of DMEs through the selective activation of PXR, CAR, and AhR.

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Abbreviations

AhR:

Aryl hydrocarbon receptor

BB:

Bilobalide

CAR:

Constitutive androstane receptor

CITCO:

6-(4-Chlorophenyl)imidazo[2,1-b][1,3]thiazole-5-carbaldehyde-O-(3,4-dichloro-benzyl)- oxime

CYP:

Cytochrome P450

GBE:

Ginkgo biloba extract

GA:

Ginkgolide A

gtPBREM:

UGT1A1 phenobarbital-responsive enhancer module

GB:

Ginkgolide B

Kae:

Kaempferol

NR:

Nuclear receptor

PXR:

Pregnane X receptor

UGT:

UDP-glucuronosyltranferases

RIF:

Rifampicin

3-MC:

3-methylcholanthrene

Que:

Quercetin

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Acknowledgments

The authors are grateful to Dr. Masahiko Negishi (National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC) for kindly providing the pCR3-hCAR, and UGT1A1-gtPBREM vectors; Dr. Curtis Omiecinski (Pennsylvania State University, University Park, Pennsylvania) for the CMV2-hCAR expression vector; and Dr. Steve Kliewer (University of Texas, Southwestern Medical Center, Dallas, TX) for the pSG5-hPXR expression vector. Human liver tissues were procured with the assistance of John Cottrell from the University of Maryland Medical Center (Baltimore, MD). This research was supported by National Institute of Health Grant (R01, DK061652).

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Li, L., Stanton, J.D., Tolson, A.H. et al. Bioactive Terpenoids and Flavonoids from Ginkgo Biloba Extract Induce the Expression of Hepatic Drug-Metabolizing Enzymes Through Pregnane X Receptor, Constitutive Androstane Receptor, and Aryl hydrocarbon Receptor-Mediated Pathways. Pharm Res 26, 872–882 (2009). https://doi.org/10.1007/s11095-008-9788-8

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