Maternal chronic unpredictable mild stress (CUMS) is associated with neuropsychiatric disorders in offspring, including anxiety, depression, and autism spectrum disorders. There is mounting evidence that these behavioral phenotypes have origins in utero, which provided that corticosterone, as a stress hormone, penetrates the placental barrier and enters the fetal body and reprograms the early neural development. Notch signaling pathway is involved in the entire life cycle from embryonic development to birth to death, and mainly regulates neural stem cell proliferation and differentiation, synaptic plasticity and neuronal damage repair, and it has been intensively involved in emotional functioning. However, the role of Notch signaling pathway in affective behaviors of offspring has not been determined. In the present study, the function of Notch signaling pathway in affective behaviors was investigated in open field test (OFT) and sucrose preference test (SPT) in offspring. The results showed that maternal chronic unpredictable mild stress alters affective behaviors in offspring. We examined HPA axis related hormones, hippocampal neuronal cell apoptosis and Notch signaling pathway in offspring. Maternal CUMS damage hippocampal pathology structure and synaptic plasticity in childhood and adolescence offspring, suggesting Notch signaling pathway in the hippocampus was activated. Furthermore, we explored the role and mechanism of Notch signaling pathway in HT22 cell damage after high corticosterone exposure in vitro. Taken together, these results indicate that maternal chronic unpredictable mild stress alters affective behaviors in offspring rats through persistent activation of Notch signaling pathway in the hippocampus.