title: Glycogen synthase kinase 3 involvement in the excessive proinflammatory response to LPS in patients with decompensated cirrhosis.
abstract: OBJECTIVE
  In decompensated cirrhosis, the early innate immune response to the Toll-like receptor 4 (TLR4) agonist, lipopolysaccharides (LPS), is characterized by a hyper-production of pro-inflammatory cytokines and hypo-production of the anti-inflammatory cytokine IL-10. In LPS-stimulated non-cirrhotic immune cells, the constitutively active glycogen synthase kinase (GSK) 3 favors pro- vs. anti-inflammatory cytokines, by acting on gene induction. However, in these cells, TLR4 dampens its own pro-inflammatory response by inducing early (within minutes) AKT-mediated phosphorylation of GSK3β (one of two GSK3 isoforms) on Ser9. Phosphorylation of GSK3β (Ser9) inhibits its activity, decreases pro-inflammatory cytokines, and increases IL-10. Thus, we investigated the role of GSK3 in LPS-induced cytokine production by peripheral blood mononuclear cells (PBMCs) or monocytes from patients with advanced cirrhosis and normal subjects.
  
  
  METHODS
  Cells were pre-incubated with or without GSK3 inhibitor (SB216763 or lithium chloride) for 1h and then stimulated with LPS. Cytokine production was assessed at mRNA and secreted proteins levels, by real-time RT-PCR at 1h and ELISA at 20 h, respectively. GSK3β phosphorylation was assessed using Western blotting.
  
  
  RESULTS
  In cirrhotic and normal PBMCs pretreated with GSK3 inhibitors, LPS-induced production of pro-inflammatory proteins TNF-α and IL-12p40 was significantly decreased while that of IL-10 was increased. LPS-induced, AKT-mediated phosphorylation of GSK3β on Ser9 found in normal monocytes, was abolished in cirrhotic cells.
  
  
  CONCLUSIONS
  GSK3 is involved in the early TLR4-mediated pro-inflammatory response in patients with decompensated cirrhosis. This was associated with a defect in AKT-mediated GSK3β phosphorylation resulting in unrestricted 'pro-inflammatory' activity of the enzyme.
authors.affiliation: INSERM, U773, Centre de Recherche Biomédicale Bichat-Beaujon CRB3, Paris and Clichy, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Coant
authors.forename: Marika
authors.initials: M
authors.lastname: Simon-Rudler
authors.forename: Thierry
authors.initials: T
authors.lastname: Gustot
authors.forename: Magali
authors.initials: M
authors.lastname: Fasseu
authors.forename: Sonia
authors.initials: S
authors.lastname: Gandoura
authors.forename: Kévin
authors.initials: K
authors.lastname: Ragot
authors.forename: Waël
authors.initials: W
authors.lastname: Abdel-Razek
authors.forename: Dominique
authors.initials: D
authors.lastname: Thabut
authors.forename: Philippe
authors.initials: P
authors.lastname: Lettéron
authors.forename: Eric
authors.initials: E
authors.lastname: Ogier-Denis
authors.forename: Romy
authors.initials: R
authors.lastname: Ouziel
authors.forename: Jacques
authors.initials: J
authors.lastname: Devière
authors.forename: Gérard
authors.initials: G
authors.lastname: Lizard
authors.forename: Zéra
authors.initials: Z
authors.lastname: Tellier
authors.forename: Didier
authors.initials: D
authors.lastname: Lebrec
authors.forename: Richard
authors.initials: R
authors.lastname: Moreau
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000328:Adult; D000368:Aged; D002478:Cells, Cultured; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000276:immunology; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D000071679:Glycogen Synthase Kinase 3 beta; D006505:Hepatitis / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D016753:Interleukin-10 / Q000276:immunology / Q000378:metabolism; D018664:Interleukin-12 / Q000276:immunology / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D018021:Lithium Chloride / Q000494:pharmacology; D008103:Liver Cirrhosis / Q000276:immunology* / Q000378:metabolism; D008297:Male; D008301:Maleimides / Q000494:pharmacology; D008875:Middle Aged; D009000:Monocytes / Q000276:immunology / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics; D012720:Severity of Illness Index; D015398:Signal Transduction / Q000276:immunology*; D051197:Toll-Like Receptor 4 / Q000276:immunology / Q000378:metabolism
chemical_list: D000276:Adjuvants, Immunologic; C508609:IL10 protein, human; D007211:Indoles; D008070:Lipopolysaccharides; D008301:Maleimides; C417521:SB 216763; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4; D016753:Interleukin-10; D018664:Interleukin-12; C494918:AKT1 protein, human; C494921:AKT2 protein, human; C491754:AKT3 protein, human; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; D051057:Proto-Oncogene Proteins c-akt; D038362:Glycogen Synthase Kinase 3; C448021:glycogen synthase kinase 3 alpha; D018021:Lithium Chloride