Brief Article
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World J Gastroenterol. May 7, 2011; 17(17): 2199-2205
Published online May 7, 2011. doi: 10.3748/wjg.v17.i17.2199
Hepatotropic growth factors protect hepatocytes during inflammation by upregulation of antioxidative systems
Matthias Glanemann, Daniel Knobeloch, Sabrina Ehnert, Mihaela Culmes, Claudine Seeliger, Daniel Seehofer, Andreas K Nussler
Matthias Glanemann, Daniel Knobeloch, Daniel Seehofer, Department of General-, Visceral- and Transplantation Surgery, Charité, Campus Virchow Klinikum, Universitätsmedizin Berlin, 13353 Berlin, Germany
Sabrina Ehnert, Mihaela Culmes, Claudine Seeliger, Andreas K Nussler, Department of Traumatology, Klinikum rechts der Isar, Technische Universität München, 81675 Munich, Germany
Author contributions: Glanemann M, Knobeloch D, Ehnert S and Nussler AK contributed equally to this work; Glanemann M, Knobeloch D, Ehnert S and Nussler AK designed the research and performed the experiments, analyzed the data and wrote the paper; Seehofer D performed the shift analysis; Culmes M and Seeliger C helped with data analysis and wrote parts of the paper.
Supported by The Federal Ministry of Research (BMBF - 01GN0984)
Correspondence to: Dr. Andreas K Nussler, Professor, Department of Traumatology, Klinikum rechts der Isar, Technische Universität München, 81675 Munich, Germany. andreas.nuessler@googlemail.com
Telephone: +49-89-41406310 Fax: +49-89-41406313
Received: July 12, 2010
Revised: August 16, 2010
Accepted: August 23, 2010
Published online: May 7, 2011
Abstract

AIM: 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 NO2-, NO3- 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 NO2-, NO3- and S-nitrosothiols in hepatocytes stimulated with LPS-containing cytokine mixture. Thus, we propose that, together with an increase in glutathione increased NO2-, NO3- formation might shift their metabolism towards non-toxic products.

CONCLUSION: 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.

Keywords: Primary human hepatocytes, Hepatocyte proliferation, Cytokines, Hepatotropic growth factors, Nitric oxide, Glutathione