Ginseng has been widely used for the treatment of various human body disorders. Despite extensive studies, the mechanism of action of ginseng remains to be clarified. In an effort of clarifying the mechanism of gluco-regulatory activity in ginseng, the effects of ginsenosides on glucose metabolism were studied in human hepatoma HepG2 cells. In this study, we showed that Rg1 significantly inhibited glucose production, glycogen synthesis, and lipogenesis. Rg1 also increased the phosphorylation of AKT and AMP-activated protein kinase (AMPK). Further studies indicated that the Rg1-mediated inhibition of glycogen and lipid synthesis was blocked in the presence of PI3K/AKT- and AMPK-specific inhibitors. These results indicate that Rg1 contributes to glucose and lipid metabolism in liver cells through AKT and AMPK pathways. Our findings provide evidence to support that Rg1 may be potential useful in counteracting insulin resistance and diabetes.