Alzheimer’s disease, a chronic and progressive neurodegenerative disorder with no prevention and cure, affecting nearly 50 million people worldwide. Glutamate is the principal excitatory neurotransmitter in the central nervous system involved in 50% of basic brain functions, especially cortical and hippocampal regions, like memory, cognition, and learning. The glutamate-mediated toxicity is termed as excitotoxicity. The present study was aimed to determine whether the methanolic and water extracts of root from the medicinal plant, Withania somnifera, could decrease the glutamate excitotoxicity and its related neuronal cell loss in a scopolamine-induced animal model of Alzheimer's disease. The rats were randomly divided into different groups of 5 in each: normal control - treated orally with saline; AD model - injected intra peritoneally with scopolamine (2 mg/Kg body wt) alone to induce Alzheimer's disease; AD model rats treated orally with the methanolic extract (AD+ME-WS) (300 mg/Kg body wt), water extract (AD+WE-WS) (300 mg/Kg body wt), and donepezil hydrochloride, a standard control (AD+DZ) (5 mg/Kg body wt) for 30 consecutive days. Increased glutamate (Glu) levels and decreased glutamate dehydrogenase (GDH) activity were reversed with Withania somnifera root extracts in both the cerebral cortex and hippocampus regions in scopolamine-induced Alzheimer's disease model rat brain. The histopathological studies of the same treatment also showed protection against neuronal cell loss in both regions. These results support the idea that these extracts could be effective for the reduction of brain damage by preventing glutamate excitotoxicity generated neuronal cell loss in the scopolamine-induced Alzheimer's disease model.