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Caspase 3 Inactivation Protects Against Hepatic Cell Death and Ameliorates Fibrogenesis in a Diet-Induced NASH Model

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Abstract

Background/Aims

Hepatocyte cell death is a key feature of nonalcoholic steatohepatitis (NASH). As the contribution of specific caspases remains unclear, our aim was to ascertain the effect of caspase 3 suppression on liver injury and fibrogenesis.

Methods

C57BL/6 wild-type (WT) and caspase 3 knock out (Casp3 −/−) mice were placed on a methionine- and choline-deficient (MCD) diet for 6 weeks to induce steatohepatitis and liver fibrosis. Thereafter, liver injury, liver fibrosis and hepatocellular apoptosis were quantified in liver sections. Additionally, expression of proteins associated with liver inflammation and fibrogenesis was analyzed.

Results

WT mice fed MCD diet showed marked activation of caspase 3 in hepatocytes, in conjunction with steatohepatitis and increased hepatic triglyceride levels, hepatocyte ballooning, inflammation and fibrosis. Casp3 −/− mice fed the MCD diet showed similar serum aminotransferase levels and NAFLD activity scores (NAS) compared with WT MCD-fed mice. However, Casp3 −/− mice on the MCD diet showed a marked reduction in expression of transcripts for profibrogenic genes, which translated into reduced hepatic collagen deposition. These changes were associated with decreased levels of apoptosis, and a significant reduction in the expression of cytokines involved in inflammatory signaling. Casp3 −/− mice on the MCD showed a reduction in expression of chemokine receptor 2 (CCR2) leading to ameliorated infiltration of inflammatory lymphocyte antigen 6 complex, locus C1 (Ly6c) positive monocytes.

Conclusion

These findings support a prominent role for hepatocyte caspase 3 activation in NASH-related apoptosis, fibrogenesis and fibrosis which in part is mediated via CCR2-dependent infiltration of Ly6c positive monocytes.

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Acknowledgments

This work was supported by NIH Grants (DK076852) and (DK082451) to AEF.

Conflict of interest

None.

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Authors and Affiliations

  1. Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA

    Samjhana Thapaliya, Michael Berk & Laura Dixon

  2. Department of Pediatrics, University of California, San Diego, 9500 Gilman Drive, MC 0715, La Jolla, CA, 92037-0715, USA

    Alexander Wree, Davide Povero, Maria Eugenia Inzaugarat & Ariel E. Feldstein

  3. Department of Pathology, University of California, San Diego, La Jolla, CA, USA

    Bettina G. Papouchado

Authors
  1. Samjhana Thapaliya
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  2. Alexander Wree
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  3. Davide Povero
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  4. Maria Eugenia Inzaugarat
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  5. Michael Berk
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  6. Laura Dixon
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  7. Bettina G. Papouchado
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  8. Ariel E. Feldstein
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Corresponding author

Correspondence to Ariel E. Feldstein.

Additional information

Samjhana Thapaliya and Alexander Wree have contributed equally to this work.

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Thapaliya, S., Wree, A., Povero, D. et al. Caspase 3 Inactivation Protects Against Hepatic Cell Death and Ameliorates Fibrogenesis in a Diet-Induced NASH Model. Dig Dis Sci 59, 1197–1206 (2014). https://doi.org/10.1007/s10620-014-3167-6

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  • DOI: https://doi.org/10.1007/s10620-014-3167-6

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