Objective: To observe the changes of depression-like behavior in SD rats induced by maternal separation (MS) stress and to explore the effects of probiotics on antidepressant-like behavior and cAMP/CREB signaling pathway. Methods: Newborn SD rats were selected as experimental subjects and divided into MS+NS group, CON+NS group, MS+P group and CON+P group using random number table method, 12 rats in each group. At PND 22-49, the MS+P and CON+P groups were given 1×109 CFU (0.1 ml) of probiotic colonies by gavage daily and the corresponding dose (1 ml/100g) according to the change of body weight, and the MS+NS and CON+NS groups were given the corresponding saline dose (1 ml/100g). Behavioral tests were performed at PND50-56, and rats were executed at PND57 for laboratory tests. Results: FST increased, OPT increased and SPT decreased after probiotic intervention. It was suggested that the MS-induced depression-like behavior was improved to some extent. Compared with the model group, probiotic intervention increased the number of neurons in the CA1 region of the hippocampus, decreased serum-associated inflammatory factors, increased serum 5-HT concentration, and decreased CORT concentration in rats. In addition, the intervention increased the expression levels of cAMP, CREB and BDNF in the hippocampus of MS rats. Conclusion: Probiotics can alleviate anxiety/depression-like behavior in SD rats, which may be related to the activation of cAMP/CREB signaling pathway. The protective effect of probiotics as therapeutic food care in preventing or alleviating MS-induced depression-like behaviors provides an experimental basis for the application of probiotics to alleviate or improve anxiety/depression.