OBJECTIVES: This study was performed to assess the effects of long-term intake of a very high carbohydrate (VHCHO) diet (76% of total energy from carbohydrate [CHO]) on whole-body glucose tolerance and hepatic insulin resistance. METHODS AND MATERIALS: Male Sprague Dawley rats were fed either a control high-CHO diet (59% total energy from CHO; n = 8) or a VHCHO diet (76% total energy from CHO; n = 8) for 17 wk. At 4, 8, 12, and 16 wk of the dietary intervention, oral glucose tolerance test and homeostasis model assessment of insulin resistance (HOMA-IR) measurements were taken to assess whole-body glucose tolerance and hepatic insulin resistance, respectively. The triacylglycerol concentration in the liver was measured at the end of the 17-wk intervention period. RESULTS: The VHCHO diet group showed significantly higher muscle glucose transporter 4 content and a smaller area under the curve for plasma glucose, but not insulin, in the oral glucose tolerance test compared with the control group. On the other hand, the VHCHO diet group had a significantly higher hepatic triacylglycerol concentration and HOMA-IR measurement compared with the control group. The hepatic triacylglycerol concentration was significantly and positively correlated with HOMA-IR. CONCLUSIONS: The results of the present study suggest that long-term intake of a VHCHO diet exerts differential effects on whole-body glucose tolerance and hepatic insulin resistance.