title: Naofen promotes TNF-α-mediated apoptosis of hepatocytes by activating caspase-3 in lipopolysaccharide-treated rats.
abstract: OBJECTIVE
  To investigate whether naofen is involved in tumor necrosis factor (TNF)-α-mediated apoptosis of hepatocytes induced by lipopolysaccharide (LPS).
  
  
  METHODS
  In vivo, rats were treated with LPS or anti-TNF-α antibody, whereas in vitro, primary hepatocytes and Kupffer cells (KCs) were separately isolated from rat livers using collagenase perfusion, and primary hepatocytes were cultured in medium containing LPS or TNF-α, or in conditioned medium from LPS-treated KCs (KC-CM)/KC-CM + anti-TNF-α antibody. Naofen and TNF-α mRNA expression was examined by real-time reverse transcription-polymerase chain reaction. Immunoblotting was used to measure protein expression. Hepatocyte apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay.
  
  
  RESULTS
  LPS significantly induced both naofen expression and caspase-3 activity in the rat liver, which coincided with an increase in the number of TUNEL-positive hepatocytes. The increase of TNF-α expression induced by LPS was preceded by increases in naofen and caspase-3 activity. Elevation of naofen expression and caspase-3 activity was abrogated by pretreatment with anti-TNF-α antibody. In KCs, LPS caused an increase in TNF-α that was almost consistent with that in the liver of LPS-treated rats. In hepatocytes, neither LPS nor TNF-α alone affected either naofen expression or caspase-3 activation. The incubation of hepatocytes with KC-CM significantly enhanced both naofen expression and caspase-3 activity. Moreover, the effects of the KC-CM-induced increase in naofen expression and caspase-3 activity were blocked by anti-TNF-α antibody.
  
  
  CONCLUSIONS
  TNF-α released from KCs treated with LPS may induce hepatic naofen expression, which then stimulates hepatocellular apoptosis through activation of caspase-3.
authors.affiliation: Jun-Hua Fan, Guo-Duo Tang, Hai-Xing Jiang, Department of Gastroenterology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China.
authors.forename: Jun-Hua
authors.initials: JH
authors.lastname: Fan
authors.affiliation: Jun-Hua Fan, Guo-Duo Tang, Hai-Xing Jiang, Department of Gastroenterology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China.
authors.forename: Guo-Gang
authors.initials: GG
authors.lastname: Feng
authors.affiliation: Jun-Hua Fan, Guo-Duo Tang, Hai-Xing Jiang, Department of Gastroenterology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Huang
authors.affiliation: Jun-Hua Fan, Guo-Duo Tang, Hai-Xing Jiang, Department of Gastroenterology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China.
authors.forename: Guo-Duo
authors.initials: GD
authors.lastname: Tang
authors.affiliation: Jun-Hua Fan, Guo-Duo Tang, Hai-Xing Jiang, Department of Gastroenterology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China.
authors.forename: Hai-Xing
authors.initials: HX
authors.lastname: Jiang
authors.affiliation: Jun-Hua Fan, Guo-Duo Tang, Hai-Xing Jiang, Department of Gastroenterology, the First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Xu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism*; D002478:Cells, Cultured; D004789:Enzyme Activation; D022781:Hepatocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D007728:Kupffer Cells / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008297:Male; D019899:Paracrine Communication / Q000187:drug effects; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism*
chemical_list: D008070:Lipopolysaccharides; D011506:Proteins; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; C582790:WDR35 protein, rat; C505410:Casp3 protein, rat; D053148:Caspase 3
keywords: Apoptosis; Caspase-3; Kupffer cells; Lipopolysaccharide; Naofen; Tumor necrosis factor-α