ObjectiveTo investigate the possible mechanism of the improvement of type 2 diabetes mellitus with insulin resistance of skeletal muscles after Roux-en-Y gastric bypass surgery (RYGB). MethodsThirty GK rats were randomly divided into GK-RYGB group, sham operation group (GK-SO group), and control group (GK-control group); in addition, 10 Wistar rats served as normal control group.On day 28, the animals were sacrificed.The ghrelin concen-tration and PI3Kp85α, Akt/PKB, and GLUT4 levels were measured by ELISA, Western blot, and real-time PCR me-thods, respectively. Results①Compared with the GK-SO group and GK-control group, the plasma ghrelin levels were significantly increased in the normal control group (P < 0.01) and GK-RYGB group (P < 0.01).②Compared with the GK-SO group and GK-control group, p-/t-PI3Kp85α, p-/t-Akt/PKB, and m-/t-GLUT4 proteins were significantly incre-ased in the normal control group (P < 0.01, P < 0.05, and P < 0.01, respectively) and GK-RYGB group (P < 0.01, P < 0.05, and P < 0.01, respectively).③Compared with the GK-SO group and GK-control group, PI3Kp85α, Akt, and GLUT4 mRNA were significantly increased in the normal control group (P < 0.01, P < 0.05, and P < 0.05, respectively) and GK-RYGB group (P < 0.01, P < 0.05, and P < 0.05, respectively). ConclusionRYGB could elevate the ghrelin level obviously and upregulate PI3Kp85α, Akt/PKB, and GLUT4 levels and thus improve the insulin resistance of skeletal muscles of rats with T2DM.