The nuclear receptor REV-ERBα regulates CYP2E1 expression and acetaminophen hepatotoxicity

Abstract

1. CYP2E1 plays an important role in drug metabolism and drug-induced hepatotoxicity. Here, we aimed to investigate a potential role for the nuclear receptor REV-ERBα in regulation of CYP2E1 expression and acetaminophen (APAP)-induced hepatotoxicity, and to determine the underlying mechanisms.

2. Regulatory effects of REV-ERBα on CYP2E1 expression were assessed in vivo (using Rev-erbα^-/- mice) and in vitro (using AML12 and HepG2 cells). In vitro microsomal CYP2E1 activity was probed using its specific substrate p-nitrophenol. Pharmacokinetic and acute toxicity studies were performed with Rev-erbα^−/− and wild-type mice after APAP administration.

3. We found that Rev-erbα ablation led to decreases in hepatic CYP2E1 expression and activity in mice. In line with this, APAP-induced hepatotoxicity was attenuated in Rev-erbα-deficient mice. The attenuated toxicity was due to down-regulation of APAP metabolism mediated by CYP2E1, which was evidenced by a decrease in formation of the toxic intermediate metabolite NAPQI (i.e. reduced APAP-cysteine and APAP-N-acetylcysteine levels). Furthermore, positive regulation of CYP2E1 expression by REV-ERBα was confirmed in both AML12 and HepG2 cells. Based on luciferase reporter assays, it was found that REV-ERBα regulated Cyp2e1 transcription and expression through repression of DEC2.

4. In conclusion, REV-ERBα positively regulates CYP2E1 expression in mice, thereby affecting APAP metabolism and hepatotoxicity.

Keywords:

• REV-ERBα
• APAP
• CYP2E1
• DEC2
• hepatotoxicity

Ethical approval

The animal study was reviewed and approved by Institutional Animal Care and Use Committee of Guangzhou University of Chinese Medicine and were performed in accordance with the NIH Guide for the Care and Use of Laboratory Animals.

Author contributions

Participated in research design: LZ, MC and BW.

Conducted experiments: LZ, FZ, YX, MC and JD.

Performed data analysis: LZ, FZ, XZ, MC and BW.

Wrote or contributed to the writing of the manuscript: LZ, MC and BW.

Disclosure statement

No potential conflict of interest was reported by the authors.

Data availability statement

The original contributions presented in the study are included in the article, further inquiries can be directed to the corresponding authors.

Additional information

Funding

This work was supported by the Natural Science Foundation of Guangdong Province [No. 2017A03031387] and the Guangzhou Science and Technology Project [No.201904010472].