Objective To observe the effects of Wuling powder and Hypericum perforatum L. on changes of behavior, the hypothalamus-pituitary-adrenal (HPA) axis function, and monoamine neurotransmitters concentration of hippocampus on chronic unpredictable stress model in rats. Methods Depression model was induced by chronic unpredictable stress combining with solitary method. The stimulus lasted for 4 weeks. During the stress, the rats were intragastric given with the decoction of Wuling powder and Hypericum perforatum L. (490 mg/kg, 245 mg/kg, 123 mg/kg) and Fluoxetine hydrochloride (2.4 mg/kg) for 4 weeks. At the end of the experiment, sucrose consumption and Morris Water Maze experiment monitor the behavior of rats, such as sucrose consumption, escape latency. Moreover, the levels of the monoamine neurotransmitters in the hippocampus were analyzed by high performance liquid chromatography-electrochemistry and the content of CORT, ACTH, CRH in plasma were detected by Radioimmunoassay. Results In Morris Water Maze test, the escape latency of rats which were consecutively administrated with Wuling powder and Hypericum perforatum L. for 4 weeks decreased obviously; the total time and distance in target quadrant remarkably increased (P < 0.05). In sucrose consumption experiment, the percentage in sugar water consumption with treatment of Wuling powder and Hypericum perforatum L. increased significantly (P < 0.01). Meanwhile, the content of norepinephrine (NE), dopamine (DA), and 5-hydroxytryptamine (5-HT) in the hippocampus were significantly increased in rats of Wuling powder and Hypericum perforatum L. and the concentrations of CORT, ACTH, CRH in plasma of Wuling powder and Hypericum perforatum L. group rats were decreased obviously (P <0.05). Conclusions The disturbances of cognition, learning and memory function of rats induced by chronic unpredictable stress combining with solitary could alleviated by Wuling powder and Hypericum perforatum L., and the pharmacological effects may related to modulation of monoaminergic neurotransmitters in the hippocampus and the content of CORT, ACTH, CRH in plasma.