Background: Smoking tobacco is a serious global health problem that is associated with psychiatric disorders and increased mortality rates. Nicotine, the main compound consumed during smoking, causes damage to different organs, especially the brain. The current study assessed the modulatory impact of Teucrium polium extract (TPE) on nicotine-induced behavioral and biochemical, alterations in mice brains. Methods: Twenty-four mice were divided into four groups and treated for three weeks. Group one was control, group two was subcutaneously injected with 2.5 mg/kg nicotine, group three received 100 mg/kg TPE orally, and group four was given 2.5 mg/kg of nicotine subcutaneously after an hour of oral administration of 100 mg/kg TPE. Result: The HPLC results of the plant extract showed the presence of 11 bioactive compounds. Nicotine administration increased anxiety and decreased locomotor activity and forelimb grip strength. Dopamine, serotonin, and acetylcholinesterase activity showed a significant decline in the nicotine-induced group. Treatment with TPE showed anxiolytic effects, modulated muscle strength and locomotor activity. Also, there was an increase in neurotransmitters compared with the nicotine group. In conclusion, TPE protected against nicotine-induced neurotoxicity through modulating behavior and neurotransmitter levels in mice.