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NLRP3 inflammasome activation is required for fibrosis development in NAFLD

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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.

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Alpha smooth muscle actin


Alanine aminotransferase


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


Arginase 1


Alcoholic steatohepatitis


Body mass index


Caspase 1


Choline-deficient amino acid-defined


Collagen, type I, alpha 1


Choline-supplemented amino acid-defined


Connective tissue growth factor


Chemokine (C-X-C motif) ligand 2


Damage associated molecular patterns


Murine macrophage marker


Hepatic stellate cell


Intercellular adhesion molecule 1




Inducible form of nitric oxide synthase


Lymphocyte antigen 6 complex


Monocyte chemotactic protein-1


Matrix metalloproteinase-2




Nonalcoholic fatty liver disease


Nonalcoholic steatohepatitis


Normal chow


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


Pathogen associated molecular patterns


Phosphate-buffered saline


Tissue inhibitor of matrix metalloproteinase 1


Tumor necrosis factor alpha


Wild type


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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).

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Correspondence to Ariel E. Feldstein.

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Wree, A., McGeough, M.D., Peña, C.A. et al. NLRP3 inflammasome activation is required for fibrosis development in NAFLD. J Mol Med 92, 1069–1082 (2014).

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