Abstract: The crude polysaccharides from Inonotus obliquus were obtained by water extraction and alcohol precipitation. And the crude polysaccharides were separated and purified from DEAE-52 cellulose column to obtain two kinds of polysaccharides(EIOP1 and EIOP2). A-Glucosidase inhibitory activity,glucose consumption of normal HepG2 cells,and insulin-resistant HepG2 cells were used as the main indicators to investigate the hypoglycemic activity of different concentrations of two purified polysaccharides from Inonotus obliquus(10,20,40,80,160,and 320 μg/mL). α-glucosidase inhibitory activity used acarbose as a positive control,and glucose consumption experiment was used with metformin as a positive control. The results showed that the α-glucosidase inhibitory activity of EIOP1 and EIOP2 were higher than that of the positive control group acarbose and crude polysaccharide,and the IC₅₀ values were 39.18 and 29.87 μg/mL,respectively. The concentration of EIOP1 on HepG2 cells was extramly significant higher than that of the control group at a concentration of 80 μg/mL(P<0.01). The promotion effect was the best,which was 30.62% higher than that of the control group. When the concentration of EIOP2 was 40 μg/mL,the glucose consumption was extramly significant higher than that of the control group,which was 75.99% higher than the control(P<0.01). The glucose consumption experiment of IR-HepG2 cells showed that EIOP1 had the best promotion effect at the concentration of 40 μg/mL,which was 30.00% higher than that of the IR group. The concentration of EIOP2 was the best at the concentration of 80 μg/mL. The IR group increased by 90.49%,which was higher than that of the Met group(P<0.01). Therefore,the purified polysaccharide of Inonotus obliquus would had a promoting effect on glucose consumption in normal HepG2 cells and insulin-resistant HepG2 cells,and better inhibitory activity against α-glucosidase than the crude polysaccharides.