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Interleukin-4 deficiency protects mice from acetaminophen-induced liver injury and inflammation by prevention of glutathione depletion

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Abstract

Objective

Interleukin-4 (IL-4) is a multifunctional cytokine involved in many diseases such as autoimmune hepatitis and idiosyncratic drug reactions. However, its role in acetaminophen (APAP)-induced liver injury remains unclear. Our objective was to evaluate the contribution of IL-4 to the pathogenesis of APAP-induced liver injury.

Methods

Balb/C (WT) and IL-4 knockout (IL-4−/−) mice were orally overdosed with APAP. After 24 h, survival percentage, biochemical and morphological markers of liver injury, and tissue inflammation were assessed.

Results

IL-4−/− mice were protected from APAP toxicity. Intravital confocal microscopy, tissue histology and serum ALT levels showed significantly less liver injury and inflammation than in the WT group, which may explain the increased survival rate of IL-4−/− mice. In addition, IL-4−/− mice had decreased production of tumor necrosis factor α, CXCL1 and interleukin-1β in the liver, but not in a remote site such as the lungs. Hepatic macrophage activation was markedly reduced in IL-4-deficient mice. In addition, glutathione depletion—a primary cause of APAP-mediated injury—was significantly attenuated in IL-4−/− mice.

Conclusions

Taken together, our data demonstrate that IL-4−/− mice are protected from APAP-induced liver injury due to reduced depletion of glutathione, which prevented liver damage and tissue inflammation.

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Acknowledgments

This study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do estado de Minas Gerais (FAPEMIG/PRONEX), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

Conflict of interest

The authors do not have any conflict of interests to disclose.

Author information

Correspondence to Gustavo Batista Menezes.

Additional information

D. A. Pires and P. E. Marques contributed equally.

Responsible Editor: Bernhard Gibbs.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Online Resource 2. 3D reconstruction of a liver from a control mouse undergoing confocal intravital microscopy. The hepatic vasculature is labeled by FITC-albumin (green) and the liver cells by Rhodamine 6G (red) (MPG 1546 kb)

Online Resource 3. 3D reconstruction of a liver from an APAP-overdosed wild-type mouse undergoing confocal intravital microscopy. The hepatic vasculature is labeled by FITC-albumin (green) and the liver cells by Rhodamine 6G (red) (MPG 1490 kb)

Online Resource 4. 3D reconstruction of a liver from an APAP-overdosed IL-4 knockout mouse undergoing confocal intravital microscopy. The hepatic vasculature is labeled by FITC-albumin (green) and the liver cells by Rhodamine 6G (red) (MPG 1434 kb)

Online Resource 1. Representative agarose gel electrophoresis of fragments generated in PCR for genotyping of wild-type (WT) and IL-4 knockout (IL-4−/−) mice. The WT mice are confirmed by the yield of a 175-basepair band and IL-4 knockout mice by a 464-basepair band in PCR (TIFF 6867 kb)

Online Resource 2. 3D reconstruction of a liver from a control mouse undergoing confocal intravital microscopy. The hepatic vasculature is labeled by FITC-albumin (green) and the liver cells by Rhodamine 6G (red) (MPG 1546 kb)

Online Resource 3. 3D reconstruction of a liver from an APAP-overdosed wild-type mouse undergoing confocal intravital microscopy. The hepatic vasculature is labeled by FITC-albumin (green) and the liver cells by Rhodamine 6G (red) (MPG 1490 kb)

Online Resource 4. 3D reconstruction of a liver from an APAP-overdosed IL-4 knockout mouse undergoing confocal intravital microscopy. The hepatic vasculature is labeled by FITC-albumin (green) and the liver cells by Rhodamine 6G (red) (MPG 1434 kb)

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Pires, D.A., Marques, P.E., Pereira, R.V. et al. Interleukin-4 deficiency protects mice from acetaminophen-induced liver injury and inflammation by prevention of glutathione depletion. Inflamm. Res. 63, 61–69 (2014) doi:10.1007/s00011-013-0671-7

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Keywords

  • Interleukin-4
  • Liver injury
  • Acetaminophen
  • Glutathione