Objective To observe the influence of Creb protein over-expression and under-expression in the prefrontal cortex on depressive behavior in rats. Methods Adeno-associated virus (AAV) strains that can knock down Creb expression in the prefrontal cortex of rats were injected using the stereotaxic injection method and screened by Western blot, reverse transcription quantitative polymerase chain reaction (RT-qPCR) and immunofluorescence. Forty rats were divided randomly into Control, chronic restraint stress (CRS), CRS combined with AAV interference (CRS+AAVI), and CRS combined with AAV overexpression (CRS+AAVO) groups. The body weight and food intake of the rats in each group were monitored during establishment of the animal model. After establishment of the model, behavioral changes in the rats were monitored by sucrose preference, elevated plus maze, forced swimming, and open field tests. The 5 hydroxytryptamine (5-HT), norepinephrine (NE), and corticosterone (CORT) contents in the prefrontal cortex of rats in each group were measured by enzyme-linked immunosorbent assay. Results Western blot and immunofluorescence showed that Creb protein expression was significantly reduced in the short hairpin RNA2 (shRNA2) knockdown groups compared with the other groups (P<0.05). RT-qPCR showed that Creb mRNA expression was also significantly reduced compared with the other three groups (P<0.01). The AAV-CREB1-shRNA2 virus strain was therefore selected for subsequent Creb knockdown experiments in this study. After modeling, the food intake of rats in the CRS+AAVI group was significantly reduced compared with the other groups (P<0.01). Rats in this group also showed slow weight gain and decreased desire to explore new environments, significantly increased despair and nervous behavior, and significantly decreased 5-HT and NE levels(P<0.01) and significantly increased CORT levels in the prefrontal cortex (P<0.01). These depressive behaviors and associated neurotransmitter levels were reversed in the CRS+AAVO group. Conclusions Lower expression of Creb in the prefrontal cortex can aggravate the degree of depression in rats, while high expression of Creb can alleviate depression to a certain extent. These result confirm that Creb expression in the prefrontal cortex is an important target in the pathogenesis of depression, thus providing ideas and references for the construction of animal gene models and further studies of the pathogenesis of depression.