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



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.


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.


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).


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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Fig. 1
Fig. 2
Fig. 3



Alanine aminotransferase


Aspartate aminotransferase





HOMA index:

Homeostasis model assessment index


Inducible nitric oxide synthase


Myeloid differentiation primary response gene 88


Non-alcoholic fatty liver disease


Non-alcoholic fatty liver disease activity score


Non-alcoholic steatohepatitis


Plasminogen activator inhibitor 1




Toll-like receptor


Tumor necrosis factor alpha


Western-style diet


Zonula occludens 1




4-Hydroxynonenal protein adducts


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

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Correspondence to Ina Bergheim.

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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.

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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).

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