gms | German Medical Science

127. Kongress der Deutschen Gesellschaft für Chirurgie

Deutsche Gesellschaft für Chirurgie

20.04. - 23.04.2010, Berlin

Establishment and characterisation of an in vitro model to research ischemia/reperfusion (I/R) injury in renal and neuronal cell culture

Meeting Abstract

  • Evelyn Jäger - Uniklinik Ulm, Thorax- und Gefäßchirurgie, Ulm, Deutschland
  • Florian Simon - Uniklinik Ulm, Thorax- und Gefäßchirurgie, Ulm, Deutschland
  • Enrico Calzia - Uniklinik Ulm, Anästhesiologische Pathophysiologie und Verfahrensentwicklung, Ulm, Deutschland
  • Peter Rademacher - Uniklinik Ulm, Anästhesiologische Pathophysiologie und Verfahrensentwicklung, Ulm, Deutschland
  • Ludger Sunder-Plassmann - Universität Ulm, Thorax- und Gefäßchirurgie, Ulm, Deutschland
  • Hubert Schelzig - Universität Ulm, Thorax- und Gefäßchirurgie, Ulm, Deutschland

Deutsche Gesellschaft für Chirurgie. 127. Kongress der Deutschen Gesellschaft für Chirurgie. Berlin, 20.-23.04.2010. Düsseldorf: German Medical Science GMS Publishing House; 2010. Doc10dgch534

DOI: 10.3205/10dgch534, URN: urn:nbn:de:0183-10dgch5348

Veröffentlicht: 17. Mai 2010

© 2010 Jäger et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielfältigt, verbreitet und öffentlich zugänglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Gliederung

Text

Introduction: Thoracic aortic clamping for vascular replacement causes eminently ischemia/ reperfusion (I/R) injury especially of the kidneys and spinal cord. Up to now there is no appropriate in vitro-model to review molecular processes during induced I/R-injury. Aim of the study is the characterisation of different cell lines during induced hypoxia/reoxidation on a molecular level. We determined a positive effect of different substances, e.g. erythropoietine (EPO) on kidney and spinal cord function during clamping/declamping of thoracic aorta in animal experiments.

Materials and methods: Analyses are arranged with human (HEK293) and porcine (LLC-PK1) cell lines. Induction of hypoxia is accomplished by regulation of oxygen concentration in a hypoxia chamber (StemCell Technologies). Among other things monitoring of apoptotic and necrotic processes as soon as the expression of different genes, e.g. hypoxia inducible factor 1 (HIF1) is carried out. Oxidative stress initially leads to an inhibition of mitochondrial oxidative phosphorylation which directly causes depletion of cellular energy sources. Therefore oxygraphical studies to ascertain mitochondrial respiration are conducted. Furthermore cells are treated with substances (EPO; carbamylated EPO, cEPO) that showed a positive effect on renal function in animal experiments.

Results: Previous data show a high tolerance of cell cultures across short times (1–12h) of hypoxia, but clear effects e.g. morphological changes and different gene expression, e.g. Bcl-2 and Bax can be seen after 24 hours of hypoxia, while damages by reoxidation could not be detected. In addition, we found significant different expression of, e.g. Bax and Bcl-2 to the point of control level after treatment with EPO and cEPO during hypoxia.

Conclusion: Establishment of a cell culture based model to investigate molecular processes during hypoxia/reoxidation according to the clinical problem of I/R is realizable. We give a reviwe of progress in our new hypoxia model.