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International Journal of Hepatology
Volume 2012, Article ID 785786, 14 pages
http://dx.doi.org/10.1155/2012/785786
Research Article

Combined Stimulation with the Tumor Necrosis Factor and the Epidermal Growth Factor Promotes the Proliferation of Hepatocytes in Rat Liver Cultured Slices

1Covance Laboratory SAS, 2-8 rue de Rouen, Z.I. de Limay-Porcheville, 78440 Porcheville, France
2Laboratoire d'Hématologie, Hôpital Pontchaillou, 2 rue Henri Le Guilloux, 35033 Rennes Cedex, France
3Inserm UMR-S 991, Université de Rennes 1, Hôpital Pontchaillou, 35033 Rennes Cedex, France
4Noscira SA, 52 Avendia de la Industria 38760 Tres Cantos, Spain

Received 1 June 2012; Revised 6 September 2012; Accepted 7 September 2012

Academic Editor: Anne Corlu

Copyright © 2012 Francis Finot et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The culture liver slices are mainly used to investigate drug metabolism and xenobiotic-mediated liver injuries while apoptosis and proliferation remain unexplored in this culture model. Here, we show a transient increase in LDH release and caspase activities indicating an ischemic injury during the slicing procedure. Then, caspase activities decrease and remain low in cultured slices demonstrating a low level of apoptosis. The slicing procedure is also associated with the G0/G1 transition of hepatocytes demonstrated by the activation of stress and proliferation signalling pathways including the ERK1/2 and JNK1/2/3 MAPKinases and the transient upregulation of c-fos. The cells further progress up to mid-G1 phase as indicated by the sequential induction of c-myc and p53 mRNA levels after the slicing procedure and at 24 h of culture, respectively. The stimulation by epidermal growth factor induces the ERK1/2 phosphorylation but fails to activate expression of late G1 and S phase markers such as cyclin D1 and Cdk1 indicating that hepatocytes are arrested in mid-G1 phase of the cell cycle. However, we found that combined stimulation by the proinflammatory cytokine tumor necrosis factor α and the epidermal growth factor promotes the commitment to DNA replication as observed in vivo during the liver regeneration.