title: Protective effects of necrostatin-1 against concanavalin A-induced acute hepatic injury in mice.
abstract: OBJECTIVE
  Necrostatin-1 (Nec-1) inhibits receptor-interacting protein 1 (RIP1) kinase and programmed necrosis. This study was designed to examine the protective effects and mechanisms of Nec-1 in concanavalin A- (ConA-) induced hepatitis in mice.
  
  
  METHODS
  C57BL/6 mice were exposed to ConA via tail vein injection and injected intraperitoneally with Nec-1 or vehicle. Levels of serum liver enzymes and histopathology were determined. Levels of inflammatory cytokines with ConA-induced hepatitis were determined with real-time polymerase chain reaction (real-time PCR). The expression of TNF- α , RIP1, and LC3 was detected with immunohistochemical staining. The expression of TNF- α , IFN- γ , IL2, IL6, caspase 3, RIP1, beclin-1, and LC3 protein was assessed by immunofluorescence and western blotting. Autophagosomes were observed with transmission electron microscopy (TEM).
  
  
  RESULTS
  Amelioration in liver functions and histopathological changes and the suppression of inflammatory cytokine production were observed in Nec-1-injected mice. Western blotting analysis showed that the expression of TNF- α , IFN- γ , IL2, IL6, and RIP1 was significantly reduced in the Nec-1-injected mice, which was confirmed by immunofluorescence and immunohistochemistry. Autophagosome formation was significantly reduced by Nec-1 treatment, as the expression of beclin-1 and LC3, determined with immunofluorescence and western blotting.
  
  
  CONCLUSIONS
  These results demonstrate that Nec-1 prevents ConA-induced liver injury via RIP1-related and autophagy-related pathways.
authors.affiliation: Department of Gastroenterology, The Tenth People's Hospital of Tongji University, Shanghai 200072, China.
authors.forename: Yingqun
authors.initials: Y
authors.lastname: Zhou
authors.forename: Weiqi
authors.initials: W
authors.lastname: Dai
authors.forename: Chunlei
authors.initials: C
authors.lastname: Lin
authors.forename: Fan
authors.initials: F
authors.lastname: Wang
authors.forename: Lei
authors.initials: L
authors.lastname: He
authors.forename: Miao
authors.initials: M
authors.lastname: Shen
authors.forename: Ping
authors.initials: P
authors.lastname: Chen
authors.forename: Chenfen
authors.initials: C
authors.lastname: Wang
authors.forename: Jie
authors.initials: J
authors.lastname: Lu
authors.forename: Ling
authors.initials: L
authors.lastname: Xu
authors.forename: Xuanfu
authors.initials: X
authors.lastname: Xu
authors.forename: Chuanyong
authors.initials: C
authors.lastname: Guo
mesh_terms: D000818:Animals; D001343:Autophagy; D056486:Chemical and Drug Induced Liver Injury / Q000188:drug therapy*; D003208:Concanavalin A / Q000633:toxicity*; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D020690:GTPase-Activating Proteins / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D007093:Imidazoles / Q000627:therapeutic use*; D007211:Indoles / Q000627:therapeutic use*; D008099:Liver / Q000473:pathology; D017114:Liver Failure, Acute / Q000188:drug therapy* / Q000517:prevention & control*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D046529:Microscopy, Electron, Transmission; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009336:Necrosis / Q000473:pathology; D010588:Phagosomes / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D000637:Transaminases / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D016207:Cytokines; D020690:GTPase-Activating Proteins; D007093:Imidazoles; D007211:Indoles; C480860:Map1lc3b protein, mouse; D008869:Microtubule-Associated Proteins; C490620:Ralbp1 protein, mouse; D014409:Tumor Necrosis Factor-alpha; C507699:necrostatin-1; D003208:Concanavalin A; D000637:Transaminases