Background: Jiawei Wendan decoction can elevate hippocampal brain-derived neurotrophic factor (BDNF) protein expression in rats with depression. It has been hypothesized that Jiawei Wendan decoction can exhibit antidepressant effects through the hippocampal signal transduction pathway of cyclic adenosine monophosphate response element binding protein (CREB)-BDNF. Objective: Using phosphorylated-CREB (p-CREB) as an entry point, the present study was designed to observe intervention effects of Jiawei Wendan decoction compared with fluoxetine. Design, time and setting: A randomized, controlled, cellular biology experiment was performed at the Central Laboratory of Guangxi University of Traditional Chinese Medicine. Materials: A total of 40 healthy, male, Sprague-Dawley rats were included in the present study. Rhizoma Acori Talarinowii (Shichangpu), Flos Albiziae (Hehuanhua), Rhizoma Pinelliae (Banxia), Caulis Bambusae in Taeniam (Zhuru), Fructus Aurantii Immaturus (Zhishi), Poria (Fuling), and Radix Bupleuri (Chaihu), the primary ingredients of Jiawei Wendan decoction, were purchased from First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine. The raw drug was decocted at a concentration of 1.5 g/mL. Fluoxetine capsules were purchased from Shanghai Zhongxi Pharmaceutical Co., Ltd., China. Methods: Following behavioral testing, 36 rats were selected from the initial 40 rats according to similar behavioral scores, and were randomly divided into 4 groups: model (n = 8), Jiawei Wendan decoction-treated (n = 10), fluoxetine-treated (n = 10), and normal control (n = 8). All rats, except for those in the normal control group, were separately raised in a chronic and unpredictable, mild-stimulation environment for 21 days to establish a depression model. The Jiawei Wendan decoction-treated and fluoxetine-treated groups were intragastrically administered Jiawei Wendan decoction (12 g/kg/d) and fluoxetine (1.8 mg/kg/d), respectively. The model and normal control groups received double-distilled water (2 mL/kg/d) once a day for a total of 21 days. Main outcome measures: Prior to depression induction and subsequent to administration, memory and degree of depression were measured by open field and tail suspension tests, respectively. At 24 hours after administration, p-CREB expression in the hippocampal CA3 region was analyzed by the immunohistochemical SABC method. Results: Two rats from the Jiawei Wendan decoction-treated group, and one from the fluoxetine-treated group, died of unknown causes. The remaining rats in each group, except for one rat that was randomly selected from the fluoxetine-treated group, were included in the final analysis. Following administration, the model group exhibited significantly decreased numbers of hippocampal p-CREB-positive cells, horizontal and vertical movements, and significantly prolonged duration of immobility in tail-suspension test, compared with the normal control group (P < 0.01). The numbers of p-CREB-positive cells, as well as horizontal and vertical movements, were significantly greater, and the duration of immobility was significantly shorter, in the Jiawei Wendan decoction and fluoxetine groups, compared with the model group (P < 0.05-0.01). There was no significant difference in the above-mentioned three indices between the Jiawei Wendan decoction and fluoxetine groups (P > 0.05). Conclusion: Jiawei Wendan decoction increased hippocampal p-CREB expression by influencing the rat hippocampal signal transduction pathway of CREB-BDNF. Jiawei Wendan decoction exhibited antidepressant effects equivalent to fluoxetine.