Abstract
Acetaminophen (APAP) overdose is a leading cause of drug-induced acute liver failure. Prolonged c-Jun N-terminal kinase (JNK) activation plays a central role in APAP-induced liver injury; however, growth arrest and DNA damage-inducible 45 beta (GADD45β) is known to inhibit JNK phosphorylation. The orphan nuclear receptor small heterodimer partner (SHP, NR0B2) acts as a transcriptional co-repressor of various genes. The aim of the present study was to investigate the role of SHP in APAP-evoked hepatotoxicity. We used lethal (750 mg/kg) or sublethal (300 mg/kg) doses of APAP-treated wild-type (WT), Shp knockout (Shp−/−), hepatocyte-specific Shp knockout (Shphep−/−), and Shp and Gadd45β double knockout (Shp−/−Gadd45β−/−) mice for in vivo studies. Primary mouse hepatocytes were used for a comparative in vitro study. SHP deficiency protected against APAP toxicity with an increased survival rate, decreased liver damage, and inhibition of prolonged hepatic JNK phosphorylation in mice, which was independent of APAP metabolism regulation. Furthermore, Shphep−/− mice showed diminished APAP hepatotoxicity compared with WT mice. SHP-deficient primary mouse hepatocytes also showed decreased cell death and inhibition of sustained JNK phosphorylation following toxic APAP treatment. While SHP expression declined, GADD45β expression increased after APAP treatment in WT mice. In Shp−/− mice, APAP-evoked GADD45β induction was significantly enhanced. Notably, the ameliorative effects of SHP deficiency on APAP-induced liver injury were abolished in Shp−/−Gadd45β−/− mice. The current study is the first to demonstrate that hepatocyte-specific SHP deficiency protects against APAP overdose-evoked hepatotoxicity in a JNK signaling regulation and GADD45β dependent manner. SHP is suggested to be a novel therapeutic target for APAP overdose treatment.
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Abbreviations
- APAP:
-
Acetaminophen
- ALT:
-
Alanine aminotransferase
- CAR:
-
Constitutive androstane receptor
- JNK:
-
c-Jun-N-terminal kinase
- CYP:
-
Cytochrome P450
- Shp flox/flox :
-
Floxed Shp exon 1 and 2
- Gadd45β −/− :
-
Gadd45β knockout
- GADD45:
-
Growth arrest and DNA damage-inducible 45
- GSH:
-
Glutathione
- H&E:
-
Hematoxylin and eosin
- MAP:
-
Mitogen-activated protein
- MKK4:
-
Kinase kinase 4
- NAQPI:
-
N-acetyl-ρ-benzoquinone imine
- NR:
-
Nuclear receptor
- ROS:
-
Reactive oxygen species
- Shp −/− Gadd45β −/− :
-
Shp and Gadd45β−/− double knockout
- Shp hep−/− :
-
Shp specifically deleted in the hepatocytes
- SHP:
-
Small heterodimer partner
- TG:
-
Transgenic
- TUNEL:
-
Terminal transferase dUTP nick end labeling
- WT:
-
Wild-type
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Acknowledgements
We thank IB Lee, YK Choi, YJ Seo and JH Choi for technical assistance. This work was supported by a grant from the National Research Foundation of Korea (NRF) and the Korean government (MSIP) (2016R1A2A1A05004858), KRIBB Research Initiative Program of the Republic of Korea, and the Development of Platform Technology for Innovative Medical Measurements Program (No. KRISS-2018-GP2018-0018) from the Korea Research Institute of Standards and Science, and received support from French state funds through the “Agence Nationale de la Recherche” under the frame programme Investissements d’Avenir labeled ANR-10-INBS-07 PHENOMIN.
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YH Kim and CH Lee designed the experiments and the study, and drafted the manuscript. YH Kim, JR Noh, DH Choi, JH Kim, SJ Moon, and JH Choi performed experiments, and collected and analyzed data for the study. Yann H provided the Shpflox/flox mice model. JH Hwang, KS Kim, TG Lee, HS Choi, and CH Lee contributed to critical revisions of the text.
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Kim, YH., Noh, JR., Hwang, J.H. et al. Hepatocyte SHP deficiency protects mice from acetaminophen-evoked liver injury in a JNK-signaling regulation and GADD45β-dependent manner. Arch Toxicol 92, 2563–2572 (2018). https://doi.org/10.1007/s00204-018-2247-3
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DOI: https://doi.org/10.1007/s00204-018-2247-3