Acacia nilotica scientifically known as Vachellia nilotica (Fabaceae family, subfamily Mimosoideae), has long been utilised to treat a variety of conditions and shows different activities such as Anti-hypertensive, antipyretics, anti-diabetic property, antioxidant, analgesic etc. This study investigates the anxiolytic effects of A. nilotica root extract in albino rat. Two polar solvents were utilised for extracting the plant's root. i.e., ethanol and distilled water. Five groups (n=6) were administered either saline, diazepam or AN roots extract administered at 200 & 400 milligram/kg concentrations orally for 28 days. The Light-dark box and Elevated Plus Maze (EPM) were used for determining anxiety levels. The extract significantly reduced anxiety-like behaviors, with treated rats spending more time in the opened arms & light chamber of the plus maze and light box. The results of the higher dose (400 mg/kg) were similar to those of diazepam, indicating that Acacia nilotica roots could potentially hold a role in the natural therapy of anxiety disorders.