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Depression is a common mental disorder and a leading cause of disability. At its most severe, it can lead to suicide. Recently, there has been growing interest in the application of natural herbs for the prevention and treatment of depression. In this report, we found that the ginsenoside active component Rg3 has an apparent antidepressant effect. In N-methyl-d-aspartic acid (NMDA)-treated HT22 murine hippocampal neuronal cells, Rg3 recovered proliferation and inhibited apoptosis by altering the cell cycle. More interestingly, Rg3 led to apparent physiological behavior change in a chronic mild stress model as seen in forced swim, tail suspension, and sucrose preference tests. This effect was mediated by the phosphorylation of cAMP response element binding protein (CREB) and brain-derived neurotrophic factor (BDNF) signaling. This study provides direct evidence to support the antidepressant effects of ginsenoside Rg3, potentially indicating its application in the treatment of clinical depression.