Nicotine withdrawal precipitates anxiety- and depression-like behaviors associated with disruptions in neurotransmitter systems, alterations in glial and neurotrophic factors, and heightened oxidative stress. This study aimed to investigate the potential therapeutic effects of magnesium sulfate (MgSO₄) on nicotine withdrawal-induced anxiety and depression in rats as well as its implications for regenerative psychiatry. These findings indicated that MgSO₄ administration effectively attenuated anxiety and depression-like behaviors induced by nicotine withdrawal in the rat model. The beneficial effects of MgSO₄ were accompanied by the modulation of cortical serotonin metabolism, a reduction in monoamine oxidase A activity, the upregulation of brain-derived neurotrophic factor, the downregulation of glial fibrillary acidic protein, and the amelioration of oxidative stress in rats with nicotine withdrawal-induced anxiety- and depression-like behaviors. Therefore, MgSO₄ can ameliorate the detrimental effects induced by nicotine withdrawal at the cellular and behavioral levels and has certain implications in regenerative psychiatry, paving the way for its therapeutic application in substance abuse-related diseases.