Hepatocyte cell death, inflammation and oxidative stress constitute key pathogenic mechanisms underlying non-alcoholic fatty liver disease (NAFLD). We aimed to investigate the role of necroptosis in human and experimental NAFLD and its association with tumour necrosis factor α (TNF-α) and oxidative stress. Serum markers of necrosis, liver receptor-interacting protein 3 (RIP3) and phosphorylated mixed lineage kinase domain-like (MLKL) were evaluated in control individuals and patients with NAFLD. C57BL/6 wild-type (WT) or RIP3-deficient (RIP3−/−) mice were fed a high-fat choline-deficient (HFCD) or methionine and choline-deficient (MCD) diet, with subsequent histological and biochemical analysis of hepatic damage. In primary murine hepatocytes, necroptosis and oxidative stress were also assessed after necrostatin-1 (Nec-1) treatment or RIP3 silencing. We show that circulating markers of necrosis and TNF-α, as well as liver RIP3 and MLKL phosphorylation were increased in NAFLD. Likewise, RIP3 and MLKL protein levels and TNF-α expression were increased in the liver of HFCD and MCD diet-fed mice. Moreover, RIP3 and MLKL sequestration in the insoluble protein fraction of NASH (non-alcoholic steatohepatitis) mice liver lysates represented an early event during stetatohepatitis progression. Functional studies in primary murine hepatocytes established the association between TNF-α-induced RIP3 expression, activation of necroptosis and oxidative stress. Strikingly, RIP3 deficiency attenuated MCD diet-induced liver injury, steatosis, inflammation, fibrosis and oxidative stress. In conclusion, necroptosis is increased in the liver of NAFLD patients and in experimental models of NASH. Further, TNF-α triggers RIP3-dependent oxidative stress during hepatocyte necroptosis. As such, targeting necroptosis appears to arrest or at least impair NAFLD progression.
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Research Article|
July 22 2015
Necroptosis is a key pathogenic event in human and experimental murine models of non-alcoholic steatohepatitis
Marta B. Afonso;
Marta B. Afonso
*Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal
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Pedro M. Rodrigues;
Pedro M. Rodrigues
*Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal
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Tânia Carvalho;
Tânia Carvalho
†Histology and Comparative Pathology Laboratory, Instituto de Medicina Molecular, Av. Prof. Egas Moniz, Edificio Egas Moniz, 1649-028 Lisbon, Portugal
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Marta Caridade;
Marta Caridade
*Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal
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Paula Borralho;
Paula Borralho
‡ESTEsL, Av. D. João II, 1990-096 Lisbon, Portugal
§Instituto de Anatomia Patológica, Universidade de Lisboa, Av. Prof. Egas Moniz, 1649-035 Lisbon, Portugal
║Hospital Cuf Descobertas, R. Mário Botas, Parque das Nações, 1998-018 Lisbon, Portugal
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Helena Cortez-Pinto;
Helena Cortez-Pinto
¶Department of Gastroenterology, Hospital Santa Maria, Faculty of Medicine, Universidade de Lisboa, Av. Prof. Egas Moniz, 1649-035 Lisbon, Portugal
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Rui E. Castro;
Rui E. Castro
*Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal
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Cecília M.P. Rodrigues
*Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisbon, Portugal
Correspondence: Cecília M.P. Rodrigues (email cmprodrigues@ff.ulisboa.pt).
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Publisher: Portland Press Ltd
Received:
November 07 2014
Revision Received:
May 22 2015
Accepted:
June 15 2015
Accepted Manuscript online:
June 15 2015
Online ISSN: 1470-8736
Print ISSN: 0143-5221
© 2015 Authors; published by Portland Press Limited
2015
Clin Sci (Lond) (2015) 129 (8): 721–739.
Article history
Received:
November 07 2014
Revision Received:
May 22 2015
Accepted:
June 15 2015
Accepted Manuscript online:
June 15 2015
Citation
Marta B. Afonso, Pedro M. Rodrigues, Tânia Carvalho, Marta Caridade, Paula Borralho, Helena Cortez-Pinto, Rui E. Castro, Cecília M.P. Rodrigues; Necroptosis is a key pathogenic event in human and experimental murine models of non-alcoholic steatohepatitis. Clin Sci (Lond) 1 October 2015; 129 (8): 721–739. doi: https://doi.org/10.1042/CS20140732
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