title: Hepatotropic growth factors protect hepatocytes during inflammation by upregulation of antioxidative systems.
abstract: OBJECTIVE
  To investigate effects of hepatotropic growth factors on radical production in rat hepatocytes during sepsis.
  
  
  METHODS
  Rat hepatocytes, isolated by collagenase perfusion, were incubated with a lipopolysaccharide (LPS)-containing cytokine mixture of interleukin-1β, tumor necrosis factor-α and interferon-γ to simulate sepsis and either co-incubated or pre-incubated with hepatotropic growth factors, e.g. hepatocyte growth factor, epidermal growth factor and/or transforming growth factor-α. Cells were analyzed for glutathione levels. Culture supernatants were assayed for production of reactive oxygen intermediates (ROIs) as well as NO(2) (-), NO(3) (-) and S-nitrosothiols. To determine cellular damage, release of aspartate aminotransferase (AST) into the culture medium was analyzed. Activation of nuclear factor (NF)-κB was measured by electrophoretic mobility shift assay.
  
  
  RESULTS
  Rat hepatocytes treated with the LPS-containing cytokine mixture showed a significant increase in ROI and nitrogen oxide intermediate formation. AST leakage was not significantly increased in cells treated with the LPS-containing cytokine mixture, independent of growth-factor co-stimulation. However, pretreatment with growth factors significantly reduced AST leakage and ROI formation while increasing cellular glutathione. Application of growth factors did not result in increased NF-κB activation. Pretreatment with growth factors further increased formation of NO(2) (-), NO(3) (-) and S-nitrosothiols in hepatocytes stimulated with LPS-containing cytokine mixture. Thus, we propose that, together with an increase in glutathione increased NO(2) (-), NO(3) (-) formation might shift their metabolism towards non-toxic products.
  
  
  CONCLUSIONS
  Our data suggest that hepatotropic growth factors positively influence sepsis-induced hepatocellular injury by reducing cytotoxic ROI formation via induction of the cellular protective antioxidative systems.
authors.affiliation: Department of General-, Visceral- and Transplantation Surgery, Charité, Campus Virchow Klinikum, Universitätsmedizin Berlin, 13353 Berlin, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Glanemann
authors.forename: Daniel
authors.initials: D
authors.lastname: Knobeloch
authors.forename: Sabrina
authors.initials: S
authors.lastname: Ehnert
authors.forename: Mihaela
authors.initials: M
authors.lastname: Culmes
authors.forename: Claudine
authors.initials: C
authors.lastname: Seeliger
authors.forename: Daniel
authors.initials: D
authors.lastname: Seehofer
authors.forename: Andreas K
authors.initials: AK
authors.lastname: Nussler
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005978:Glutathione / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D007249:Inflammation / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D016211:Transforming Growth Factor alpha / Q000494:pharmacology*; D015854:Up-Regulation
chemical_list: D000975:Antioxidants; D016328:NF-kappa B; D017382:Reactive Oxygen Species; D016211:Transforming Growth Factor alpha; D009569:Nitric Oxide; D004815:Epidermal Growth Factor; D017228:Hepatocyte Growth Factor; D005978:Glutathione