title: NLRP3 inflammasome activation in D-galactosamine and lipopolysaccharide-induced acute liver failure: role of heme oxygenase-1.
abstract: D-Galactosamine (GalN) and lipopolysaccharide (LPS) are commonly used to study mechanisms of hepatic malfunction that result in hepatic inflammation and subsequent fulminant hepatic failure. Inflammasomes are intracellular multiprotein complexes that in response to cellular danger signals trigger the biological maturation of proinflammatory cytokines. Heme oxygenase-1 (HO-1) is a cytoprotective enzyme that induces anti-inflammatory and antioxidant activity against oxidative cellular stress. This study examined activation of the NACHT, LRR, and PYD domain-containing protein 3 (NLRP3) inflammasome in GalN/LPS-induced hepatic injury and the role of HO-1 in the signaling pathways of inflammasome. Mice (C57BL/6) were pretreated twice with hemin (HO-1 inducer, 30 mg/kg) and zinc protoporphyrin (ZnPP; HO-1 inhibitor, 10mg/kg) at 12 and 2h before GalN (800 mg/kg)/LPS (40 μg/kg) administration. HO-1 induction with hemin reversed the lethality induced by GalN/LPS administration, and ZnPP pretreatment blocked this change. Lipid peroxidation markedly increased after GalN/LPS treatment, whereas glutathione content decreased in the GalN/LPS group. These changes were attenuated by hemin, but ZnPP reversed the effects of hemin. Serum levels of tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β increased after GalN/LPS treatment; these increases were attenuated by hemin. Hepatic mRNA levels of TNF-α, IL-1β, and NLRP3 increased after GalN/LPS treatment, and hemin attenuated increases in TNF-α and IL-1β. After GalN/LPS treatment, the hepatic expression of NLRP3, ASC, and caspase-1 (p10) was increased. In immunoprecipitation studies, hemin attenuated the interaction of NLRP3 with ASC and caspase-1. GalN/LPS induced expression of the thioredoxin-interacting protein (TXNIP) gene and the interaction between NLRP3 and TXNIP; again, hemin attenuated these effects. The effects of hemin were reversed by ZnPP. Our findings suggest that activation of the NLRP3 inflammasome leads to a GalN/LPS-induced inflammatory response through TXNIP-NLRP3 interaction. Furthermore, HO-1 overexpression may protect the liver against GalN/LPS-induced inflammation through suppression of the NLRP3 signaling pathway.
authors.affiliation: School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
authors.forename: Seok-Joo
authors.initials: SJ
authors.lastname: Kim
authors.affiliation: School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea. Electronic address: sunmee@skku.edu.
authors.forename: Sun-Mee
authors.initials: SM
authors.lastname: Lee
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D020170:Caspase 1 / Q000235:genetics / Q000378:metabolism; D005688:Galactosamine; D005978:Glutathione / Q000378:metabolism; D051547:Heme Oxygenase-1 / Q000502:physiology*; D058847:Inflammasomes / Q000502:physiology*; D053583:Interleukin-1beta / Q000235:genetics / Q000378:metabolism; D015227:Lipid Peroxidation; D008070:Lipopolysaccharides / Q000494:pharmacology; D008099:Liver / Q000201:enzymology / Q000276:immunology / Q000473:pathology; D017114:Liver Failure, Acute / Q000139:chemically induced / Q000201:enzymology* / Q000276:immunology; D008297:Male; D008565:Membrane Proteins / Q000502:physiology*; D008810:Mice, Inbred C57BL; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D058847:Inflammasomes; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C492716:Nlrp3 protein, mouse; C486715:TXNIP protein, human; D014409:Tumor Necrosis Factor-alpha; D005688:Galactosamine; D051547:Heme Oxygenase-1; C496542:Hmox1 protein, mouse; D020170:Caspase 1; D005978:Glutathione
keywords: ASC; Acute liver failure; Free radicals; GSH; GSSG; GalN; HO; Heme oxygenase-1; IL; Interleukin-1β; LPS; MDA; NACHT, LRR, and pyrin domain-containing protein 3; NF-κB; NLRP3; NLRP3 inflammasome; ROS; Reactive oxygen species; SDS; TLR; TNF; TRX; TXNIP; Toll-like receptor; ZnPP; apoptosis-associated speck-like protein containing a CARD; d-galactosamine; heme oxygenase; interleukin; lipopolysaccharide; malondialdehyde; nuclear factor κB; oxidized glutathione; reactive oxygen species; reduced glutathione; sodium dodecyl sulfate; thioredoxin; thioredoxin-interacting protein; tumor necrosis factor; zinc protoporphyrin