A Combination of Leucine, Metformin, and Sildenafil Treats Nonalcoholic Fatty Liver Disease and Steatohepatitis in Mice
Sild-Met-Leu treatment in hepatocytes increases AMPK and Sirt1 activation and stimulates fatty acid oxidation. HepG2 cells were treated with sildenafil (Sild, 1 nM), metformin (Met, 0.1 mM), and leucine (Leu, 0.5 mM) as indicated for 24 hours after induction of lipid accumulation and compared to nontreated cells (control or CTRL high) after lipid accumulation and without lipid accumulation (CTRL low). (a) Sirt1 and (b) AMPK and phospho-AMPK protein expression was measured via Western blotting. Representative blots are shown. Data from repeated experiments are analyzed and presented as mean ± SEM ( to 8). (c) Oxygen consumption rate (OCR) after 200 μM palmitate injection was measured and the area under the curve (AUC) was calculated. Data are represented as mean ± SEM ().