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The essential role of CYP2E1 in metabolism and hepatotoxicity of N,N-dimethylformamide using a novel Cyp2e1 knockout mouse model and a population study

  • Zhijun Wu
  • Qiang Liu
  • Lei Wang
  • Min Zheng
  • Mingyue Guan
  • Man Zhang
  • Wenjin Zhao
  • Chunmin Wang
  • Songwen Lu
  • Juan ChengEmail author
  • Shuguang LengEmail author
Toxicogenomics
  • 21 Downloads

Abstract

N,N-Dimethylformamide (DMF) is a widespread contaminant of leather factories and their surrounding environment. There is a lack of direct in vivo evidence supporting CYP2E1 as a primary enzyme responsible for DMF metabolism and hepatotoxicity. In this study, a novel Cyp2e1 knockout (KO) mouse model was generated and used to assess whether DMF metabolism and hepatotoxicity is CYP2E1 dependent using an acute toxicity protocol with a single dose of 1500 mg DMF/kg. An epidemiological study in 698 DMF-exposed workers and 188 non-DMF-exposed controls was conducted to investigate the associations between functional polymorphisms of CYP2E1 (rs6413432/rs2031920) and DMF metabolite (N-methylcarbmoylated-hemoglobin [NMHb]). We successfully established Cyp2e1 KO mice with evidence from DNA sequence analysis, which showed 1-bp insertion at 65 bp (C) site of Cyp2e1 Exon 1. In addition, western blot and in vivo pharmacokinetic study also showed a complete absence of CYP2E1 protein and a 92% and 88% reduction in CYP2E1 activity among males and females, respectively. DMF metabolism as evidenced by increased blood NMHb, and hepatotoxicity as evidenced by elevated liver/body weight ratio, activity of liver enzymes and massive liver necrosis were detected in wild-type (WT) mice but were completely abrogated in KO mice, strongly supporting a CYP2E1-dependent pattern of DMF metabolism and hepatotoxicity. Moreover, variant allele of CYP2E1-rs6413432 was also significantly associated with higher NMHb levels in DMF-exposed workers (P = 0.045). The increase of glucose-regulated protein 94 detected in WT mice but not in KO mice suggested CYP2E1-dependent endoplasmic reticulum stress may be a key mechanism underlying DMF-induced hepatotoxicity.

Keywords

CYP2E1 Knockout (KO) mouse N,N-Dimethylformamide (DMF) Hepatotoxicity Metabolism 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 81372963). We thank Ms Maria Picchi from Lovelace Respiratory Research Institute for scientific editing of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

204_2019_2567_MOESM1_ESM.doc (86 kb)
Supplementary material 1 (DOC 87 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.The Toxicology Laboratory of National Institute of Occupational Health and Poison ControlChinese Center for Disease Control and PreventionBeijingChina
  2. 2.Department of Environmental HealthSuzhou Center for Disease Control and PreventionJiangsuChina
  3. 3.Department of Physical and Chemical LaboratorySuzhou Center for Disease Control and PreventionJiangsuChina
  4. 4.School of Public HealthQingdao UniversityShandongChina

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