Our results demonstrate that chronic alcohol feeding results in a decrease in AMPK activity, which is recovered by RGE treatment. Previously, we reported that feeding mice with a Lieber–DeCarli diet containing 5% EtOH for 10 days, followed by a single dose of EtOH gavage (5 g/kg body weight) (chronic–binge EtOH model) induces significant fatty liver and liver injury

with oxidative stress (Fig. 6A) [25]. To investigate the effect of RGE for the treatment of selleck chemicals ALD using the chronic–binge EtOH model, EtOH-fed mice were treated with RGE. Treatment with RGE decreased EtOH-induced serum ALT and AST levels (Fig. 6B). The protective effect of RGE on alcoholic steatosis was further confirmed by liver histology as shown by H&E staining. It was noted that treatment of alcohol-fed mice with RGE completely inhibited fat infiltration (Fig. 6C), confirming Selleckchem Bioactive Compound Library the ability of RGE to inhibit fat accumulation in liver. Moreover, the chronic–binge EtOH model significantly increased 4-HNE positive cells, which is consistent with our previous report [25]. However, similar to the chronic EtOH model, the amount of 4-HNE positive cells was dose-dependently and significantly reduced by treatment with RGE (Fig. 7A). RGE also markedly attenuated nitrotyrosine positive cells, confirming that RGE is capable of inhibiting alcohol-induced oxidative stress in the chronic–binge EtOH animal model (Fig. 7B). We next examined the effect of RGE on

fat accumulation in a mouse hepatocyte cell line, AML12. EtOH treatment for 3 days increased fat accumulation in hepatocytes as Osimertinib datasheet shown by Oil red O staining. However, RGE (500 μg/mL or 1000 μg/mL) treatment reduced fat accumulation in a dose-dependent manner (Fig. 8A). To determine whether changes of fat accumulation in the hepatocyte were consistent with lipogenesis- or lipolytic-associated gene expression, the expression of SREBP-1, Sirt1, and PPARα was observed by Western blot analysis following concomitant treatment with 10–1000 μg/mL of RGE and EtOH for 3 days. In agreement with the in vivo data, RGE inhibited the ability of EtOH to induce SREBP-1 and repress Sirt1

and PPARα expression in AML12 cells ( Fig. 8B). The pharmacological properties of ginseng are primarily attributed to a group of active ingredients, the ginsenosides, which are a diverse group of steroidal saponins. Gum and Cho recently reported that total ginsenoside amount of RGE was 19.66 mg/g containing the major ginsenosides Rb1 (4.62 mg/g), Rb2 (1.83 mg/g), Rc (2.41 mg/g), Rd (0.89 mg/g), Re (0.93 mg/g), Rf (1.21 mg/g), Rg1 (0.71 mg/g), Rg2 (3.21 mg/g), Rg3 (3.05 mg/g), Rh1 (0.78 mg/g), and other minor ginsenosides [21]. Therefore, we next identified the major component of red ginseng required for the inhibition of hepatic steatosis. We determined the effects of the major ginsenosides Rb1, Rb2, and Rd on the EtOH-induced fat accumulation in AML12 cells.