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A dual function of the furanocoumarin chalepensin in inhibiting Cyp2a and inducing Cyp2b in mice: the protein stabilization and receptor-mediated activation

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Abstract

Chalepensin, a furanocoumarin, is present in several medicinal Rutaceae plants and causes a mechanism-based inhibition of human and mouse cytochrome P450 (P450, CYP) 2A in vitro. To address the in vivo effect, we investigated the effects of chalepensin on multiple hepatic P450 enzymes in C57BL/6JNarl mice. Oral administration of 10 mg/kg chalepensin to mice for 7 days significantly decreased hepatic coumarin 7-hydroxylation (Cyp2a) and increased 7-pentoxyresorufin O-dealkylation (Cyp2b) activities, whereas activities of Cyp1a, Cyp2c, Cyp2e1, and Cyp3a were not affected. Without affecting its mRNA level, the decreased Cyp2a activity was accompanied by an increase in the immunodetected Cyp2a5 protein level. In chalepensin-treated mice, microsomal Cyp2a5 was less susceptible to ATP-fortified cytosolic degradation than that in control mice, resulting in the elevated protein level. The in vitro inactivation through NADPH-fortified pre-incubation with chalepensin also protected microsomal Cyp2a5 against protein degradation. Using cell-based reporter systems, chalepensin at a concentration near unbound plasma concentration activated mouse constitutive androstane receptor (CAR), in agreement with the observed induction of Cyp2b. These findings revealed that suicidal inhibition of Cyp2a5 and the CAR-mediated Cyp2b9/10 induction concurrently occurred in chalepensin-treated mice.

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Abbreviations

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

CAR:

Constitutive androstane receptor

COH:

Coumarin 7-hydroxylation

CPR:

NADPH-cytochrome P450 reductase

EROD:

7-Ethoxyresorufin O-deethylation

8-MOP:

8-Methoxypsoralen

MROD:

7-Methoxyresorufin O-demethylation

P450:

Cytochrome P450

PROD:

7-Pentoxyresorufin O-dealkylation

PXR:

Pregnane X receptor

TCPOBOP:

1,4-Bis-[2-(3,5-dichloropyridyloxy)]benzene

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Acknowledgments

We sincerely appreciate the technical assistance from Miss Yu-Ping Chang (National Research Institute of Chinese Medicine, Taipei). This work was supported by grants from the National Science Council, Taipei (grant number: NSC97-2320-B-077-009-MY3), National Research Institute of Chinese Medicine, Taipei and China Medical University, Taichung, Taiwan (grant number: CMU100-N2-04).

Conflict of interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. National Research Institute of Chinese Medicine, No. 155-1, Li-Nong Street, Sec. 2, Taipei, 112, Taiwan, ROC

    Wei-Sheng Lo, Chien-Chih Chen, Chih-Chien Hsu & Yune-Fang Ueng

  2. Department of Pharmacy, College of Pharmacy, China Medical University, 91 Hsueh-Shih Rd., Taichung, 404, Taiwan, ROC

    Yun-Ping Lim & Yune-Fang Ueng

  3. Department of Emergency, Toxicology Center, China Medical University Hospital, Taichung, 404, Taiwan, ROC

    Yun-Ping Lim

  4. Department of Biotechnology, Hungkuang University, No. 34, Chung-Chie Rd., Shalu, Taichung County, 433, Taiwan, ROC

    Chien-Chih Chen

  5. Institute of Oral Biology, School of Dentistry, National Yang-Ming University, No. 155, Li-Nong Street, Sec. 2, Taipei, 112, Taiwan, ROC

    Chih-Chien Hsu & Yune-Fang Ueng

  6. Department of Toxicogenomics, National Institute of Public Health, Šrobárova 48, 10042, Praha 10, Czech Republic

    Pavel Souček

  7. School of Biological Sciences and Technology, Chonnam National University, Gwangju, 500-757, Republic of Korea

    Chul-Ho Yun

  8. Center for Pharmacogenetics, University of Pittsburgh, 633 Salk Hall, 3501 Terrace Street, Pittsburgh, PA, 15261, USA

    Wen Xie

  9. Institute of Medical Science, Taipei Medical University, No. 250, Wuxing Street, Taipei, 110, Taiwan, ROC

    Yune-Fang Ueng

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  1. Wei-Sheng Lo
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  2. Yun-Ping Lim
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  4. Chih-Chien Hsu
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  7. Wen Xie
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  8. Yune-Fang Ueng
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Correspondence to Yune-Fang Ueng.

Additional information

Wei-Sheng Lo and Yun-Ping Lim have contributed equally to this work.

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Lo, WS., Lim, YP., Chen, CC. et al. A dual function of the furanocoumarin chalepensin in inhibiting Cyp2a and inducing Cyp2b in mice: the protein stabilization and receptor-mediated activation. Arch Toxicol 86, 1927–1938 (2012). https://doi.org/10.1007/s00204-012-0902-7

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  • DOI: https://doi.org/10.1007/s00204-012-0902-7

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