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Oral citrulline supplementation protects female mice from the development of non-alcoholic fatty liver disease (NAFLD)

European Journal of Nutrition volume 56, pages 2519–2527 (2017)Cite this article

Abstract

Purpose

Impairments of intestinal barrier function are discussed as risk factors for the development and progression of non-alcoholic fatty liver disease (NAFLD). Studies suggest an association between arginine/citrulline homeostasis and the development of liver damages. Here, the effect of an oral l-citrulline (Cit) supplement on the development of a Western-style diet (WSD)-induced NAFLD was determined in mice.

Methods

Female 6- to 8-week-old C57BL/6J mice were either pair-fed a liquid Western-style or control diet (C) ± 2.5 g/kg bodyweight Cit for 6 weeks (C + Cit or WSD + Cit). Indices of liver damage, glucose metabolism, intestinal barrier function and NO synthesis were measured.

Results

While bodyweight gain was similar between groups, markers of glucose metabolism like fasting blood glucose and HOMA index and markers of liver damage like hepatic triglyceride levels, number of neutrophils and plasminogen activator inhibitor-1 protein levels were significantly lower in WSD + Cit-fed mice when compared to WSD-fed mice only. Protein levels of the tight junction proteins occludin and zonula occludens-1 in duodenum were significantly lower in mice fed a WSD when compared to those fed a WSD + Cit (−~70 and −~60 %, respectively, P < 0.05), whereas portal endotoxin levels, concentration of 3-nitrotyrosine protein adducts in duodenum and toll-like receptor-4 mRNA expression in livers of WSD + Cit-fed mice were markedly lower than in WSD-fed mice (−~43 %, P = 0.056; −~80 and −~48 %, respectively, P < 0.05).

Conclusion

Our data suggest that the protective effects of supplementing Cit on the development of NAFLD in mice are associated with a decreased translocation of endotoxin into the portal vein.

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Abbreviations

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

C:

Control

Cit:

l-Citrulline

HOMA index:

Homeostasis model assessment index

iNOS:

Inducible nitric oxide synthase

MYD88:

Myeloid differentiation primary response gene 88

NAFLD:

Non-alcoholic fatty liver disease

NAS:

Non-alcoholic fatty liver disease activity score

NASH:

Non-alcoholic steatohepatitis

PAI-1:

Plasminogen activator inhibitor 1

TG:

Triglycerides

Tlr:

Toll-like receptor

TNFα:

Tumor necrosis factor alpha

WSD:

Western-style diet

ZO-1:

Zonula occludens 1

3-NT:

3-Nitrotyrosine

4-HNE:

4-Hydroxynonenal protein adducts

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Acknowledgments

The present study was funded by a grant from the German Research Foundation (DFG): BE 2376/6-1 (I. B.).

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Authors and Affiliations

  1. Institute of Nutritional Sciences, SD Model Systems of Molecular Nutrition, Friedrich-Schiller University Jena, Dornburger Str. 29, 07743, Jena, Germany

    Cathrin Sellmann, Cheng Jun Jin, Anna Janina Engstler & Ina Bergheim

  2. Nutrition Biology Laboratory EA4466, Faculty of Pharmacy, Paris Descartes University, Sorbonne Paris Cité, Paris, France

    Jean-Pascal De Bandt

  3. Clinical Chemistry Department, Paris Centre University Hospitals, APHP, Paris, France

    Jean-Pascal De Bandt

Authors
  1. Cathrin Sellmann
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  2. Cheng Jun Jin
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  3. Anna Janina Engstler
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  4. Jean-Pascal De Bandt
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  5. Ina Bergheim
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Corresponding author

Correspondence to Ina Bergheim.

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Conflict of interest

C. Sellmann, C.J. Jin, A.J. Engstler and I. Bergheim have no conflicts of interest. J-P De Bandt is a shareholder of Citrage company.

Ethical approval

Approval for all experiments was provided by the local Institutional Animal Care and Use Committee (IACUC).

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Sellmann, C., Jin, C.J., Engstler, A.J. et al. Oral citrulline supplementation protects female mice from the development of non-alcoholic fatty liver disease (NAFLD). Eur J Nutr 56, 2519–2527 (2017). https://doi.org/10.1007/s00394-016-1287-9

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  • DOI: https://doi.org/10.1007/s00394-016-1287-9

Keywords

  • Citrulline
  • Intestinal barrier function
  • Endotoxin
  • Occludin
  • Non-alcoholic fatty liver disease