Hypoxia-reoxygenation-induced chemokine transcription is not prevented by preconditioning or intermittent hypoxia, in mice hepatocytes.
Prolonged ischemia used in liver surgery and/or transplantation causes cellular damage resulting in apoptosis and necrosis. Ischemia-reperfusion (I/R) led Kupffer cells to pro-inflammatory cytokines secretion [tumor necrosis factor (TNF)-alpha, interleukin-1] which involve chemokines secretion by hepatocytes. These chemokines have neutrophil chemotactic properties and neutrophils are involved in the development of I/R-induced necrosis. The aim of this study was to specify the consequence of partial oxygen pressure variation on hepatocyte chemokines synthesis and to verify if intermittent hypoxia and/or preconditioning could decrease it. It was performed on primary cultured mice hepatocytes and Kupffer cells, subjected to continuous, intermittent hypoxia or preconditioning phases, mimicking surgical processes. The chemokine secretion was evaluated by RNase protection assay and enzyme-linked immunosorbent assay method. Only monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-2 (MIP-2) mRNA formation were observed, especially after 1-h hypoxia followed by 10-h (for MCP-1) or 24-h reoxygenation (for MIP-2). In conclusion, TNF-alpha and coculture with Kupffer cells increased hepatocyte chemokines mRNA transcription, whereas surgical split up protocols (intermittent hypoxia and preconditioning) had no significant effect.
Laurens
Marina
M
Laboratoire de Recherches Chirurgicales, Faculté de Médecine, Université de Nice-Sophia Antipolis, France.
Defamie
Virginie
V
Scozzari
Gitana
G
Schmid-Alliana
Annie
A
Gugenheim
Jean
J
Crenesse
Dominique
D
D000818:Animals; D015687:Cell Hypoxia / Q000502:physiology*; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism; D054412:Chemokine CCL8; D018925:Chemokines / Q000235:genetics*; D022781:Hepatocytes / Q000187:drug effects / Q000502:physiology*; D019194:Ischemic Preconditioning; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D018945:Monocyte Chemoattractant Proteins / Q000235:genetics / Q000378:metabolism; D010100:Oxygen / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects*
C111835:CCL2 protein, human; C515453:CCL8 protein, human; C515454:Ccl8 protein, mouse; D018932:Chemokine CCL2; D054412:Chemokine CCL8; D018925:Chemokines; D018945:Monocyte Chemoattractant Proteins; D010100:Oxygen