A Combination of Leucine, Metformin, and Sildenafil Treats Nonalcoholic Fatty Liver Disease and Steatohepatitis in Mice
Sild-Met-Leu treatment decreases inflammatory marker in hepatocytes and macrophages. Mouse hepatocytes (AML cells) and macrophages (RAW 264.7 cells), grown individually or in coculture, were treated with sildenafil (1 nM), metformin (0.1 mM), and leucine (0.5 mM) as indicated for 24 hours after induction with free fatty acids (FFA) and/or LPS. Nontreated cells (control) were included for comparison. ((a) and (b)) Monocyte chemotactic protein- (MCP-) 1 secretion in media of AML cells and of hepatocyte-macrophage coculture. Data are presented as mean ± SEM ( to 10). ((c) to (e)) Protein expression of phosphorylated and total NF-κB and TNF-alpha in AML 12 hepatocytes and RAW 264.7 macrophages. Quantitative data are presented as mean ± SEM (), and representative blots are shown. (f) Macrophage tumor necrosis factor- (TNF-) alpha secretion was measured in the media. Data are presented as mean ± SEM ().