title: Hepatocytes produce TNF-α following hypoxia-reoxygenation and liver ischemia-reperfusion in a NADPH oxidase- and c-Src-dependent manner.
abstract: Cell line studies have previously demonstrated that hypoxia-reoxygenation (H/R) leads to the production of NADPH oxidase 1 and 2 (NOX1 and NOX2)-dependent reactive oxygen species (ROS) required for the activation of c-Src and NF-κB. We now extend these studies into mouse models to evaluate the contribution of hepatocytes to the NOX- and c-Src-dependent TNF-α production that follows H/R in primary hepatocytes and liver ischemia-reperfusion (I/R). In vitro, c-Src-deficient primary hepatocytes produced less ROS and TNF-α following H/R compared with controls. In vivo, c-Src-KO mice also had impaired TNF-α and NF-κB responses following partial lobar liver I/R. Studies in NOX1 and p47phox knockout primary hepatocytes demonstrated that both NOX1 and p47phox are partially required for H/R-mediated TNF-α production. To further investigate the involvement of NADPH oxidases in the production of TNF-α following liver I/R, we performed additional in vivo experiments in knockout mice deficient for NOX1, NOX2, p47phox, Rac1, and/or Rac2. Cumulatively, these results demonstrate that NOX2 and its activator subunits (p47phox and Rac) control the secretion of TNF-α by the liver following I/R. Interestingly, in the absence of Kupffer cells and NOX2, NOX1 played a dominant role in TNF-α production following hepatic I/R. However, NOX1 deletion alone had little effect on I/R-induced TNF-α. Thus Kupffer cell-derived factors and NOX2 act to suppress hepatic NOX1-dependent TNF-α production. We conclude that c-Src and NADPH oxidase components are necessary for redox-mediated production of TNF-α following liver I/R and that hepatocytes play an important role in this process.
authors: Department of Anatomy and Cell Biology, The University of Iowa, Iowa City, IA 52242, USA.
authors: Netanya Y
authors: NY
authors: Spencer
authors: Weihong
authors: W
authors: Zhou
authors: Qiang
authors: Q
authors: Li
authors: Yulong
authors: Y
authors: Zhang
authors: Meihui
authors: M
authors: Luo
authors: Ziying
authors: Z
authors: Yan
authors: Thomas J
authors: TJ
authors: Lynch
authors: Duane
authors: D
authors: Abbott
authors: Botond
authors: B
authors: Banfi
authors: John F
authors: JF
authors: Engelhardt
mesh_terms: D000818:Animals; D015153:Blotting, Western; D000081247:CSK Tyrosine-Protein Kinase; D005682:Gadolinium; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D022781:Hepatocytes / Q000378:metabolism*; D008099:Liver / Q000098:blood supply / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D018345:Mice, Knockout; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015427:Reperfusion Injury / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism*; D019061:src-Family Kinases / Q000235:genetics / Q000378:metabolism*
chemical_list: D016328:NF-kappa B; D017382:Reactive Oxygen Species; D014409:Tumor Necrosis Factor-alpha; D005682:Gadolinium; D019255:NADPH Oxidases; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; C038958:gadolinium chloride
keywords: NADPH oxidase; c-Src; hepatocyte; redox; reperfusion injury
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