title: Suppression of autophagy sensitizes Kupffer cells to endotoxin.
abstract: OBJECTIVE
  Recent evidence suggests that protein degradation system autophagy is implicated in a component of innate immunity. We report here that suppression of autophagy in Kupffer cells due to hepatic steatosis enhances an inflammatory response to endotoxin.
  
  
  METHODS
  Kupffer cells were isolated from C57BL/6J mice fed chow diet (control) or high-fat diet (HFD) for 12 weeks, liver-specific autophagy-deficient mice (Atg7(F/F) :Mx1-Cre) and wild-type mice (Atg7(F/F) ). Kupffer cells were incubated with 100 ng/mL lipopolysaccharide (LPS). The concentration of tumor necrosis factor (TNF)-α in media was measured by enzyme-linked immunoassay. Expression of Toll-like receptor (TLR)4, IκB kinase (IKK)-α/β, p38, p62 and LC3 in Kupffer cells was evaluated by western blot analysis.
  
  
  RESULTS
  Incubation with LPS increased LC3-II expression of Kupffer cells from control mice; however, an increase in LC3-II expression due to LPS was suppressed in Kupffer cells from HFD mice. Moreover, both p62 expression and TNF-α production in Kupffer cells from HFD mice was higher than control mice. On the other hand, LPS exposure increased TNF-α production from autophagy-deficient Kupffer cells more than wild type. There was no significant difference in expression of TLR4 between wild and autophagy-deficient Kupffer cells. Nevertheless, activation of p38 or IKK in Kupffer cells due to LPS was augmented by autophagy deficiency. The addition of the p38 inhibitor SB203580 attenuated TNF-α production in both wild and autophagy-deficient Kupffer cells.
  
  
  CONCLUSIONS
  These results suggest that suppression of autophagy observed in Kupffer cells from steatotic liver sensitizes to endotoxin. In conclusion, suppression of autophagy may play a pivotal role on progression of NAFLD.
authors.affiliation: Department of Gastroenterology, Juntendo University School of Medicine             Protein Metabolism Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
authors.forename: Hiroo
authors.initials: H
authors.lastname: Fukada
authors.forename: Shunhei
authors.initials: S
authors.lastname: Yamashina
authors.forename: Kousuke
authors.initials: K
authors.lastname: Izumi
authors.forename: Masaaki
authors.initials: M
authors.lastname: Komatsu
authors.forename: Keiji
authors.initials: K
authors.lastname: Tanaka
authors.forename: Kenichi
authors.initials: K
authors.lastname: Ikejima
authors.forename: Sumio
authors.initials: S
authors.lastname: Watanabe