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

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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.

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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

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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.

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Authors and Affiliations

  1. Division of Gastroenterology, Department of Medicine, Duke University Medical Center, 905 LaSalle Street, Snyderman Building, Suite 1073, Durham, NC, 27710, USA

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

  2. Gastroenterology Department, Hospital de Santa Maria, CHLN, Lisbon, Portugal

    Mariana Verdelho Machado

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  1. Mariana Verdelho Machado
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  2. Leandi Kruger
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  4. Gregory Alexander Michelotti
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  8. Anna Mae Diehl
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Correspondence to Anna Mae Diehl.

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Machado, M.V., Kruger, L., Jewell, M.L. et al. Vitamin B5 and N-Acetylcysteine in Nonalcoholic Steatohepatitis: A Preclinical Study in a Dietary Mouse Model. Dig Dis Sci 61, 137–148 (2016). https://doi.org/10.1007/s10620-015-3871-x

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