To observe the effects of transcutaneous auricular vagus nerve stimulation (taVNS) on plasma melatonin (MLT) content and insulin receptor expression in the liver, the skeletal muscles, and the pancreas of Zucker diabetic fatty (ZDF) rats, so as to explore the hypoglycemic mechanism of taVNS. Thirty male ZDF rats were randomly divided into model group, taVNS group and sham-taVNS group, with 10 rats in each group; besides, 10 male Zucker lean rats of the same strain were collected for the blank control group. ZDF rats were fed with high-fat diet to induce type 2 diabetes mellitus (T2DM) rat model. In the taVNS group, HANS-100A electroacupuncture instrument was used to stimulate the cavum conchae of both sides. The stimulation sites of rats in the sham-taVNS were the same as the taVNS group, but without electricity delivered. The above interventions were performed 30 min each time, once daily, lasting for 6 weeks. Fasting blood glucose (FBG) was measured weekly in each group, the plasma metatonin (MLT) content was detected by ELISA, and the insulin receptor expression level in the liver, the skeletal muscle and the pancreas was determined by Western blot. Compared with the blank control group, the level of FBG of rats were increased (P<0.01), the plasma MLT content was decreased (P<0.01) and the insulin receptor expression level in the pancreatic tissue was decreased (P<0.01) in the model group. In the taVNS gruop, FBG was decreased (P<0.05, P<0.01), the plasma MLT content was increased (P<0.01), and the insulin receptor expression level in the liver, the skeletal muscle and the pancreas was increased (P<0.05, P<0.01, P<0.001) when compared with the model group. Compared with the taVNS group, FBG was increased (P<0.05, P<0.01), the plasma MLT content was decreased (P<0.01), and the expression level of insulin receptors in the skeletal muscle and the pancreas was decreased (P<0.01, P<0.001) in the sham-taVNS group. The taVNS can improve the insulin resistance and ultimately obtain the antihyperglycemic effect through regulating MLT concentration.