Objective: In this study， the effects of different dietary fatty acid composition on liver glucose level were discussed based on the interaction of insulin and iron metabolism. Methods: Sixty 21-day-old clean male SD rats were randomly divided into: control group， 15% SO group， 15% PO group (lard)， 15% OO group (olive oil)， 15% FO group (fish oil)， and 1∶1 SO∶FO group. Firstly， the fatty acid compositions of oils were determined by gas chromatography， and then the contents of glucose and insulin in liver were measured after 13 weeks of intervention with different oils. The contents of hemoglobin， serum iron， ferritin in liver were also detected， and the iron deposition in liver tissue was observed by prussian blue staining. The key genes expressions of iron metabolism in rat liver was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Results: The n-6 unsaturated fat acid content in SO was 46.37%， the n-3 unsaturated fat acid content in FO was 37.55%， the saturated fat acid content in PO was 24.78%， and the monounsaturated fatty acid content in OO was 36.39%. There were no significant differences in the contents of glucose in the liver of rats in each group. The level of hepatic insulin in the rats of 1∶1 SO∶FO group was significantly lower than that in the control group and the 15% SO group (P < 0.05). There were no significant differences in hemoglobin and ferritin among all the groups， but the serum iron content in 15% SO group was the highest and significantly higher than that in the control group and 1∶1 SO∶FO group (P < 0.05). Compared with the control group， the iron content in liver of 15% PO group was significantly higher (P < 0.05)， and prussian blue staining results showed that brown particles increased in 15% SO group and 15% PO group. The expression of hepcidin (Hep) gene decreased significantly in 15% SO group and 15% PO group (P < 0.05)； the expression of ferroportin 1 (FPN 1) gene in 15% SO group and 15% PO group was significantly lower than those in other high-fat groups (P < 0.05). Compared with the control group， the expression of divalent metal transporter 1 (DMT-1) gene in 15% FO group was significantly higher (P < 0.05). Compared with the control group， the expression of transferrin receptor-2 (TfR-2) gene in 15% PO group decreased significantly (P < 0.05). Conclusion: Compared with 15% SO， 15% 1∶1 SO∶FO could inhibit the increase of serum iron level and liver iron deposition， and then resist the decrease of insulin sensitivity； 15% OO and 15% FO had no remarkable effect on liver iron deposition， while 15% PO could cause liver iron deposition. Overall， appropriate supplementation of fish oil can improve liver glucose and iron metabolism.