Objective To study the changes of behavior and depression-like classic indicators after hippocampal microinjection of K252a, and to establish a new animal model of depression. Methods SD rats were randomly divided into five groups, namely the control group, sham group, chronic stress depression model group, hippocampal of K252a microinjection group, and hippocampal microinjection K252a plus chronic stress group. Open field experiments, sucrose consumption test, and Morris water maze behavioral assay were used to assess the behavioral changes in the rats. ELISA was used to detect the plasma monoamine neurotransmitter, radioimmunoassay was used to determine the plasma CRH, ACTH, CORT contents, and western-blotting was performed to observe the protein expression of BDNF, CREB, ERK1/2, and BCL-2 in the hippocampus. Results Compared with the control group, the amount of activity, sugar consumption, learning and memory abilities were decreased(P<0.05 or P<0.01), also the serum monoamine neurotransmitters were decreased (P<0.01), HPA axis function was improved (P<0.01), and the expression of BDNF, CREB, ERK1/2, BCL-2 decreased in the CUMS group(P<0.05 or P<0.01), but there was no significant difference in the DMSO group.Compared with the DMSO group, the activity, consumption of sucrose, learning and memory ability were significantly decreased(P<0.05 or P<0.01),while the HPA axis function was increased (P<0.05 or P<0.01),the serum monoamine neurotransmitters decreased(P<0.05 or P<0.01), and the BDNF, CREB, ERK1/2, BCL-2 expressions in the hypocampus were significantly decreased(P<0.05 or P<0.01) in the K252a group and K252a + CUMS group. Compared with the CUMS group,the K252a group and K252a + CUMS group did not show significant changes in these parameters. Compared with the K252a group, these indicators were not significantly changed in the K252a + CUMS group. Conclusions The results of behavior, hematology, and molecular biology analysis show that this model has a great similarity to the classical model of CUMS in surface validity, construct validity, and functional validity. It may provide an alternative investigative technology platform for basic research and antidepressant drug screening.