[51] In the context of early activation

of the inflammasome in ASH and a significant protection from all selleck inhibitor components of alcoholic liver disease observed in mice deficient in inflammasome components or IL-1 signaling, the available data suggested differential role of inflammasomes in the pathogenesis of ASH and NASH. In search for mechanisms that would explain this discrepancy, we investigated the cellular source of activated inflammasome and IL-1β. Both in ASH and NASH, the baseline levels of caspase-1 protein or pro-Casp-1, ASC, Nlrp3, and pro-IL-1b mRNA were substantially higher in liver immune cells, compared to hepatocytes.[66, 67] Specific for ASH, analysis of liver immune cells or primary hepatocytes isolated from alcohol-fed mice showed that alcohol increased the active fragment of caspase-1 and IL-1β only in liver immune cells but not in primary hepatocytes. As these data suggested that liver immune cells were the predominant cell type that activates caspase-1 and IL-1β in ASH, we generated Deforolimus cell line caspase-1-chimeric mice using a combination of clodronate-mediated Kupffer cell depletion, irradiation, and bone marrow transplantation. Using this model, we confirmed our hypothesis

that caspase-1 expressed in Kupffer cells was involved in alcohol-induced liver inflammation, steatosis, and injury, and we did not find any evidence for a pathogenic role for caspase-1 in liver parenchymal cells in the development of ASH.[67] In addition to Kupffer cell-specific

inflammasome activation in ASH,[67] we observed that activation of the inflammasome occurred also in isolated hepatocytes in NASH.[66] Specifically, primary hepatocytes isolated from the MCD-fed mice had increased expression of NALP3, ASC, caspase-1, and pro-IL-1b mRNA.[49, 66] Taking into account that fatty livers had elevated the expression of inflammasome components and that this process occurred in hepatocytes which accumulate lipids, we tested whether fatty acids exert any effects on inflammasome in hepatocytes. We observed that in vitro treatment of primary mouse hepatocytes with palmitoic acid, a MCE saturated fatty acid, resulted in upregulation of the inflammasome component NALP3, priming of caspase-1 for subsequent activation by LPS and induction of IL-1β secretion. Using the pan-caspase inhibitor Z-VAD, we demonstrated that these events were caspase dependent, and we also showed that they were caused by saturated fatty acids, whereas non-saturated fatty acids had no effect. We further showed that hepatocytes exposed to palmitoic acid produced inflammasome-mediated danger signals, which in turn activated liver macrophages in a caspase-dependent manner.[66] Taken together, our findings have outlined several differences in inflammasome/IL-1 signaling between ASH and NASH.