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Journal of Molecular Medicine

, Volume 92, Issue 10, pp 1069–1082 | Cite as

NLRP3 inflammasome activation is required for fibrosis development in NAFLD

  • Alexander Wree
  • Matthew D. McGeough
  • Carla A. Peña
  • Martin Schlattjan
  • Hongying Li
  • Maria Eugenia Inzaugarat
  • Karen Messer
  • Ali Canbay
  • Hal M. Hoffman
  • Ariel E. Feldstein
Original Article

Abstract

NLR inflammasomes, caspase 1 activation platforms critical for processing key pro-inflammatory cytokines, have been implicated in the development of nonalcoholic fatty liver disease (NAFLD). As the direct role of the NLRP3 inflammasome remains unclear, we tested effects of persistent NLRP3 activation as a contributor to NAFLD development and, in particular, as a modulator of progression from benign hepatic steatosis to steatohepatitis during diet-induced NAFLD. Gain of function tamoxifen-inducible Nlrp3 knock-in mice allowing for in vivo temporal control of NLRP3 activation and loss of function Nlrp3 knockout mice were placed on short-term choline-deficient amino acid-defined (CDAA) diet, to induce isolated hepatic steatosis or long-term CDAA exposure, to induce severe steatohepatitis and fibrosis, respectively. Expression of NLRP3 associated proteins was assessed in liver biopsies of a well-characterized group of patients with the full spectrum of NAFLD. Nlrp3 −/− mice were protected from long-term feeding CDAA-induced hepatomegaly, liver injury, and infiltration of activated macrophages. More importantly, Nlrp3 −/− mice showed marked protection from CDAA-induced liver fibrosis. After 4 weeks on CDAA diet, wild-type (WT) animals showed isolated hepatic steatosis while Nlrp3 knock-in mice showed severe liver inflammation, with increased infiltration of activated macrophages and early signs of liver fibrosis. In the liver samples of patients with NAFLD, inflammasome components were significantly increased in those patients with nonalcoholic steatohepatitis (NASH) when compared to those with non-NASH NAFLD with mRNA levels of pro-IL1 beta correlated to levels of COL1A1. Our study uncovers a crucial role for the NLRP3 inflammasome in the development of NAFLD. These findings may lead to novel therapeutic strategies aimed at halting the progression of hepatic steatosis to the more severe forms of this disease.

Key message

  • Mice with NLRP3 inflammasome loss of function are protected from diet-induced steatohepatitis.

  • NLRP3 inflammasome gain of function leads to early and severe onset of diet-induced steatohepatitis in mice.

  • Patients with severe NAFLD exhibit increased levels of NLRP3 inflammasome components and levels of pro-IL1β mRNA correlate with the expression of COL1A1.

Keywords

NLRP3 Inflammation Liver fibrosis NASH Steatoheptatitis 

Abbreviations

α-SMA

Alpha smooth muscle actin

ALT

Alanine aminotransferase

ASC

Apoptosis-associated speck-like protein containing a caspase recruitment domain

Arg1

Arginase 1

ASH

Alcoholic steatohepatitis

BMI

Body mass index

Casp1

Caspase 1

CDAA

Choline-deficient amino acid-defined

COL1A1

Collagen, type I, alpha 1

CSAA

Choline-supplemented amino acid-defined

CTGF

Connective tissue growth factor

CXCL 2

Chemokine (C-X-C motif) ligand 2

DAMPs

Damage associated molecular patterns

F4/80

Murine macrophage marker

HSC

Hepatic stellate cell

ICAM1

Intercellular adhesion molecule 1

IL

Interleukin

iNOS

Inducible form of nitric oxide synthase

Ly6c

Lymphocyte antigen 6 complex

MCP1

Monocyte chemotactic protein-1

MMP2

Matrix metalloproteinase-2

MPO

Myeloperoxidase

NAFLD

Nonalcoholic fatty liver disease

NASH

Nonalcoholic steatohepatitis

NC

Normal chow

NLRs

Nucleotide-binding oligomerization domain (NOD) leucine-rich-repeat containing receptors

PAMPs

Pathogen associated molecular patterns

PBS

Phosphate-buffered saline

TIMP1

Tissue inhibitor of matrix metalloproteinase 1

TNF-α

Tumor necrosis factor alpha

WT

Wild type

Notes

Acknowledgments

We thank Martin Pronadl and Rudolf Ott from the Clinic of Surgery II at Alfried Krupp Hospital Essen, Germany, for collecting tissue and serum samples during bariatric surgeries and clinical follow up of the enrolled patients. We thank Bettina Papouchado for assessing steatosis, inflammation, and ballooning in the liver samples. This work was funded by NIH (DK076852 and DK082451 to AEF and AI52430 to HMH) and the German Research Foundation (DFG-grant 173/2-1 to AW).

Conflict of interest

The authors state that they have nothing to disclose.

Supplementary material

109_2014_1170_MOESM1_ESM.docx (17 kb)
Supplemental Table 1 (DOCX 17 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Alexander Wree
    • 1
    • 2
  • Matthew D. McGeough
    • 1
  • Carla A. Peña
    • 1
  • Martin Schlattjan
    • 2
  • Hongying Li
    • 3
  • Maria Eugenia Inzaugarat
    • 4
  • Karen Messer
    • 3
  • Ali Canbay
    • 2
  • Hal M. Hoffman
    • 1
    • 5
  • Ariel E. Feldstein
    • 1
  1. 1.Department of PediatricsUniversity of California, San DiegoLa JollaUSA
  2. 2.Department of Gastroenterology and HepatologyUniversity Hospital EssenEssenGermany
  3. 3.Biostatistics and Bioinformatics Group, Moores Cancer CenterUniversity of California, San DiegoLa JollaUSA
  4. 4.Institute of Immunology, Genetics and MetabolismCONICET-UBABuenos AiresArgentina
  5. 5.Ludwig Institute of Cancer Research, San DiegoLa JollaUSA

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