The present study was designed to clarify the principal herb responsible for the improvement of insulin resistance produced by Die-Huang-Wan, a mixture of six herbs, in rats fed with fructose-rich chow for 4 weeks. A decrease in plasma glucose was observed in fructose-rich chow-fed rats received an oral administration of Die-Huang-Wan at 26 mg/kg for 60 min but it disappeared with the deletion of dioscorea (Dioscoreae rhizoma) while this action was not modified by the deletion of other five herbs. The decrease of plasma glucose in fructose-rich chow-fed rats produced by dioscorea was similar to that treated with Die-Huang-Wan at same dosing; while the other five herbs failed to produce same influence. Similar to the effect of Die-Huang-Wan, dioscorea improved the fructose-induced decrement of insulin-stimulated glucose disposal rate after 3 days of treatment. Also, oral administration of dioscorea at effective dose (4.2 mg/kg per administration, three times daily) into streptozotocin-induced diabetic rats for 10 days increased the response to exogenous insulin, approaching to that induced by Die-Huang-Wan in same treatment. However, these effects failed to induce in the dioscorea-deleted formula of Die-Huang-Wan or other five herbs of this mixture. These results suggest that dioscorea is the major herb for the improvement of insulin sensitivity produced by Die-Huang-Wan. This can be applied to use as an adjuvant for subjects who need to increase insulin sensitivity.