The purpose of the study is to investigate the effect of 8-hydroxy-dihydroberberine on insulin   resistance induced by high free fatty acid (FFA) and high glucose in 3T3-L1 adipocytes and its possible molecular mechanism.  Palmic acid or glucose in combination with insulin was used to induce insulin resistance in 3T3-L1 adipocytes.  8-Hydroxy-dihydroberberine and berberine were added to the cultured medium separately, which were considered as treated group and positive control group.  The rate of glucose uptake was determined by 2-deoxy-³H-D-glucose method.  The amount of glucose consumption in the medium was measured by glucose oxidase method.  Cell growth and proliferation of 3T3-L1 adipocytes were detected with Cell Counting Kit-8 (CCK-8) assay.  After incubated with palmic acid for 24 hours or glucose with insulin for 18 hours, the rate of glucose transport in 3T3-L1 adipocytes was inhibited by 67% and 58%, respectively.  The amount of glucose consumption in 3T3-L1 adipose cells was decreased by 41% after cells were incubated with palmic acid for 24 h.  However, the above changes were reversed by pretreatment with 8-hydroxy-dihydroberberine for 24 and 48 h.  Significant difference existed between groups.  Insulin resistance in 3T3-L1 adipocytes, which is induced by high FFA and high glucose, could be ameliorated by 8-hydroxy-dihydroberberine.