NLRP3 inflammasome activation is required for fibrosis development in NAFLD
- 2.1k Downloads
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.
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.
KeywordsNLRP3 Inflammation Liver fibrosis NASH Steatoheptatitis
Alpha smooth muscle actin
Apoptosis-associated speck-like protein containing a caspase recruitment domain
Body mass index
Choline-deficient amino acid-defined
Collagen, type I, alpha 1
Choline-supplemented amino acid-defined
Connective tissue growth factor
- CXCL 2
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
Nonalcoholic fatty liver disease
Nucleotide-binding oligomerization domain (NOD) leucine-rich-repeat containing receptors
Pathogen associated molecular patterns
Tissue inhibitor of matrix metalloproteinase 1
Tumor necrosis factor alpha
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.
- 16.Kamari Y, Shaish A, Vax E, Shemesh S, Kandel-Kfir M, Arbel Y, Olteanu S, Barshack I, Dotan S, Voronov E et al (2011) Lack of interleukin-1alpha or interleukin-1beta inhibits transformation of steatosis to steatohepatitis and liver fibrosis in hypercholesterolemic mice. J Hepatol 55:1086–1094PubMedCrossRefPubMedCentralGoogle Scholar
- 17.de Roos B, Rungapamestry V, Ross K, Rucklidge G, Reid M, Duncan G, Horgan G, Toomey S, Browne J, Loscher CE et al (2009) Attenuation of inflammation and cellular stress-related pathways maintains insulin sensitivity in obese type I interleukin-1 receptor knockout mice on a high-fat diet. Proteomics 9:3244–3256PubMedCrossRefGoogle Scholar
- 19.Wree A, Eguchi A, McGeough MD, Pena CA, Johnson CD, Canbay A, Hoffman HM, Feldstein AE (2013) NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation and fibrosis. Hepatology 59:898–910 Google Scholar
- 20.Kodama Y, Kisseleva T, Iwaisako K, Miura K, Taura K, De Minicis S, Osterreicher CH, Schnabl B, Seki E, Brenner DA (2009) c-Jun N-terminal kinase-1 from hematopoietic cells mediates progression from hepatic steatosis to steatohepatitis and fibrosis in mice. Gastroenterology 137:1467–1477, e1465PubMedCrossRefPubMedCentralGoogle Scholar
- 27.Kalsch J, Bechmann LP, Kalsch H, Schlattjan M, Erhard J, Gerken G, Canbay A (2011) Evaluation of biomarkers of NAFLD in a cohort of morbidly obese patients. J Nutr Metab 2011: 369168. doi: 10.1155/2011/369168
- 30.Miura K, Kodama Y, Inokuchi S, Schnabl B, Aoyama T, Ohnishi H, Olefsky JM, Brenner DA, Seki E (2010) Toll-like receptor 9 promotes steatohepatitis by induction of interleukin-1beta in mice. Gastroenterology 139(323–334):e327Google Scholar
- 43.Wree A, Bechmann LP, Claudel T, Schlattjan M, Sowa JP, Baba H, Gerken G, Feldstein AE, Trauner M, Canbay A (2013) Bariatric surgery reduces adipocyte size, improves liver injury and counteracts lipotoxicity via changes in serum fatty acid composition and adiponectin levels. J Hepatol 58:S553Google Scholar