Add time:09/25/2019         Source:sciencedirect.com

Kupffer cells are involved in phagocytosis and known to release biologically active mediators during early events of liver injury. Such functional properties of Kupffer cells can be modulated by methyl palmitate (MP). Therefore, efficacy of MP to modulate Kupffer cell function was evaluated in cultured primary Kupffer cells from rat liver. Phagocytic activity of Kupffer cells was measured by their capacity to phagocytize latex beads and the release of TNF-α, IL-10, IL-6, nitric oxide, and PGE2 was determined in cell culture medium after incubating the cells with various concentrations of MP for 24 h followed stimulation with lipopolysaccharide (LPS) for 6 h. To understand the mechanism of phagocytosis, we investigated the hydrolysis of MP, and determine ATP levels and activity of NF-κB in MP-inhibited Kupffer cells. A significant decrease was observed in phagocytosis. Phagocytosis evaluated at 0.5 mM MP was found to be time-dependent with a maximum decrease of 49% at 6 h. Exposure of Kupffer cells to MP followed by LPS stimulation showed a dose-dependent decrease in phagocytosis and reduced the release of TNF-α, IL-10, nitric oxide, and PGE2 but not of IL-6 levels in the supernatant as compared to the control. While ATP levels were unchanged, the nuclear factor NF-κB (p65) activity was inhibited in Kupffer cells treated with MP after LPS stimulation (35.6 RLU versus 49.6 RLU in control). Hydrolysis of MP was found to be time-dependent; maximum concentration of MP and palmitic acid (hydrolysis products) in the cell being at 3 and 6 h, respectively. In general, MP appears to reduce phagocytosis and levels of TNF-α, IL-10, nitric oxide, and PGE2 without affecting ATP levels and is probably mediated by NF-κB. This in vitro model is useful for detailed mechanistic studies of inhibition of phagocytosis by MP and other fatty acid esters.

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