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Digestive Diseases and Sciences

, Volume 61, Issue 1, pp 137–148 | Cite as

Vitamin B5 and N-Acetylcysteine in Nonalcoholic Steatohepatitis: A Preclinical Study in a Dietary Mouse Model

  • Mariana Verdelho Machado
  • Leandi Kruger
  • Mark L. Jewell
  • Gregory Alexander Michelotti
  • Thiago de Almeida Pereira
  • Guanhua Xie
  • Cynthia A. Moylan
  • Anna Mae Diehl
Original Article

Abstract

Background

Nonalcoholic fatty liver disease (NAFLD) is the number one cause of chronic liver disease and second indication for liver transplantation in the Western world. Effective therapy is still not available. Previously we showed a critical role for caspase-2 in the pathogenesis of nonalcoholic steatohepatitis (NASH), the potentially progressive form of NAFLD. An imbalance between free coenzyme A (CoA) and acyl-CoA ratio is known to induce caspase-2 activation.

Objectives

We aimed to evaluate CoA metabolism and the effects of supplementation with CoA precursors, pantothenate and cysteine, in mouse models of NASH.

Methods

CoA metabolism was evaluated in methionine–choline deficient (MCD) and Western diet mouse models of NASH. MCD diet-fed mice were treated with pantothenate and N-acetylcysteine or placebo to determine effects on NASH.

Results

Liver free CoA content was reduced, pantothenate kinase (PANK), the rate-limiting enzyme in the CoA biosynthesis pathway, was down-regulated, and CoA degrading enzymes were increased in mice with NASH. Decreased hepatic free CoA content was associated with increased caspase-2 activity and correlated with worse liver cell apoptosis, inflammation, and fibrosis. Treatment with pantothenate and N-acetylcysteine did not inhibit caspase-2 activation, improve NASH, normalize PANK expression, or restore free CoA levels in MCD diet-fed mice.

Conclusion

In mice with NASH, hepatic CoA metabolism is impaired, leading to decreased free CoA content, activation of caspase-2, and increased liver cell apoptosis. Dietary supplementation with CoA precursors did not restore CoA levels or improve NASH, suggesting that alternative approaches are necessary to normalize free CoA during NASH.

Keywords

Nonalcoholic steatohepatitis Coenzyme A Caspase-2 Pantothenate N-Acetylcysteine 

Abbreviations

NAFLD

Nonalcoholic fatty liver disease

NASH

Nonalcoholic steatohepatitis

CoA

Coenzyme A

MCD

Methionine–choline deficient

PANK

Pantothenate kinase

WT

Wild type

ALT

Alanine aminotransferase

AST

Aspartate aminotransferase

TBARS

Thiobarbituric acid-reactive substances

α-SMA

Alpha-smooth muscle actin

SOD

Superoxide dismutase

GPX

Glutathione peroxidase

4-HNE

4-Hydroxynonenal

TNF-α

Tumor necrosis factor alpha

Notes

Financial Support

This research is supported by NIH R01 DK077794-08, R37 AA010154-19 and R56 DK106633-01 (Diehl AM), and Duke Endowment: The Florence McAlister Professorship (Diehl AM). MVM is the recipient of a PhD grant from Fundação para a Ciência e Tecnologia, FCT, Portugal.

Compliance with ethical standards

Conflict of interest

None.

Supplementary material

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Supplementary material 1 (DOCX 58 kb)
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Supplementary material 2 (TIFF 526 kb)
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Supplementary material 3 (TIFF 5998 kb)
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Supplementary material 4 (TIFF 11122 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Mariana Verdelho Machado
    • 1
    • 2
  • Leandi Kruger
    • 1
  • Mark L. Jewell
    • 1
  • Gregory Alexander Michelotti
    • 1
  • Thiago de Almeida Pereira
    • 1
  • Guanhua Xie
    • 1
  • Cynthia A. Moylan
    • 1
  • Anna Mae Diehl
    • 1
  1. 1.Division of Gastroenterology, Department of MedicineDuke University Medical CenterDurhamUSA
  2. 2.Gastroenterology Department, Hospital de Santa MariaCHLNLisbonPortugal

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