During critical periods of brain development, exercise-induced physical fitness may greatly impact the brain structure and function. Nevertheless, forced and intensive physical activities may display negative effects, particularly in the pre-pubertal period. Preadolescent rats were exposed to an enriched environment and combined exercise training for three consecutive weeks in the present study. There was a large cage with enriching stimuli and voluntary physical activity opportunities as an enriched environment (EE). The combined exercise training (CET) consisted of aerobic and resistance training programs. The protein levels of corticosterone (CORT), glucocorticoid receptors (GRs), insulin-like growth factor-1 (IGF-1), brain-derived neurotrophic factor (BDNF), and vascular endothelial growth factor (VEGF) were assessed using Enzyme-linked immunosorbent assay and western blotting. Cresyl violet staining was also used to evaluate the number of cells in the hippocampus. While GRs levels were significantly increased in both EE and CET groups (P < 0.001), decreased CORT levels were found in enriched rats (P < 0.05). Moreover, elevated BDNF levels were found in the EE (P < 0.01) and CET (P < 0.05) groups. Similarly, VEGF significantly increased in the EE (P < 0.01) and CET (P < 0.05) animals. However, IGF-1 levels were high only in trained rats (P < 0.05). The number of cells also significantly increased in the DG and CA1 region of the hippocampus after each intervention (P < 0.001). These findings clarified that combined exercise training and voluntary physical activity in an enriched environment during the preadolescent period might promote the downstream plasticity effects on the hippocampus.