Precision-cut liver slices in culture as a tool to assess the physiological involvement of Kupffer cells in hepatic metabolism

Excerpt

Hepatic macrophages have the capacity to secrete a tremendous array of molecules, which can be divided into 3 categories é cytokines (TNF-alpha), lipid mediators (prostaglandins PGE₂) and reactive intermediates (NOé) é in response to stimulus, such as lipopolysaccharides (LPS) [1, 2]. Such mediators are capable to modulate both the metabolism and the integrity of hepatocytes in vitro [2]. The physiological role of Kupffer cell in hepatic metabolism regulation has been approached in the present study by using the original in vitro model of precision-cut liver slices (PCLS) in culture; this model allows preserving the liver lobule architecture, by maintaining namely cell diversity in physiological proportion and cell-cell interactions [3]. First, we established whether non-parenchymal cells are still viable in rat PCLS and are able to respond to LPS in vitro; TNF-alpha, PGE₂, NOx (reflecting NOé release) were measured in the incubation medium of PCLS from rats previously treated with GdCl₃ é a specific inhibitor of Kupffer cell phagocytosis [4] é or NaCl as a control- in order to evaluate the contribution of Kupffer cell in mediator release. Moreover, by using the same model, we have investigated the role of Kupffer cell in the regulation of lipid synthesis in PCLS, in order to approach the biochemical mechanism explaining our last results, which indicate that the inhibition of Kupffer cell by GdCl₃ leads to triglycerides accumulation in liver tissue [5]. …