Oxidative stress response induced in rat primary hepatocyte monolayers by mechanical removal of adherent cells |
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Authors: | T. M. Pillar Hans Joachim Seitz |
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Affiliation: | Abt. Biochem. Endokrinologie, Institut für Physiologische Chemie, Universit?ts-Krankenhaus Eppendorf, D-20246 Hamburg, Germany Tel.: +49 40 4717 3387; Fax: +49 40 4717 4862; e-mail: PILLAR@uke.uni-hamburg.de, DE
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Abstract: | We show that mechanical separation of adherent rat primary hepatocytes after the monolayer-forming stage causes the induction of the oxidative stress genes HO-1 (haem oxygenase) and MnSOD (manganese superoxide dismutase). The procedures for enzymatically breaking up liver tissue structure and isolating hepatocytes do not cause HO-1 and MnSOD activation. Only after a 3-h incubation, during which hepatocytes form a monolayer on culture dishes, does the hydrodynamic shearing away of necrotic cells sticking to the monolayer surface activate these two genes. Analysis of this injury-response pathway shows that oxidative stress and mitochondrial dysfunction play a role, as activation can be repressed by antioxidants and by respiratory inhibitors. Recovery of the cells takes a further 24-h incubation during which HO-1 and MnSOD expression returns to basal levels. |
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