Maternal Bisphenol A (BPA) diet triggers anxiety in rodents, but the underlying mechanism is still unclear. Accumulating epidemiological and experimental data have demonstrated that the anxiety is associated with aberrant neuroimmune response. In this study, we found that maternal BPA diet (MBD) exacerbated anxiety-like behavior in female juvenile mice, and the molecular evidence further showed that this behavioral phenotype was connected to the neuroimmune activation, such as elevated tumor necrosis factor alpha (TNF-α) and interleukin (IL)-6 levels in prefrontal cortex (PFC) rather than in peripheral blood, which indicated that neuroimmune response might be ascribed to neuroglial activation because activated neuroglia cells could secrete proinflammatory cytokines. Subsequently, we found that ionized calcium-binding adapter molecule (Iba)-1 as a selective marker for microglia and glial fibrillary acidic protein as a specific marker for astrocyte were significantly increased at transcriptional and translational levels, which confirmed the neuroglial activation in this model. Therefore, we conclude that MBD induces excessive anxiety-like behavior in female juvenile with elevated TNF-α and IL-6 levels, as well as activated microglia and astrocyte in PFC. Herein caution must be taken to prevent potential risks from MBD becuase exacerbated anxiety-like behavior in female juvenile by MBD may be a critical contribution for subsequent growth or mental disorders.