Stroke is the third cause of death and a leading cause of chronic disability all around the world. Inflammation and oxidative stress are common pathogenic pathways of brain injury induced by ischemia/reperfusion (I/R). This work aims to investigate the possible role of Piperine as antioxidant and anti-inflammatory in attenuating the cerebral ischemia/reperfusion (I/R)-induced brain injury in whole body γ-irradiated rats. Global cerebral ischemia was induced in both irradiated and non-irradiated male Wistar rats, and was followed by reperfusion. The beneficial effect of pretreatment with piperine (10 mg/kg, once daily p.o. for 15 days) was examined on cerebral ischemia-induced brain injury in 24 h pre-irradiated (6 Gy) male Wistar rats. I/R increased brain content of malondialdehyde (MDA) and decreased its glutathione (GSH) content and total antioxidant capacity (TAC). It also raised serum lactate dehydrogenase (LDH) activity and brain cytosolic calcium (Ca2+) level. Pre-treatment with piperine significantly attenuated brain injury in I/R irradiated rats by decreasing cytosolic brain calcium level and serum lactate dehydrogenase (LDH) activity. In addition, a significant decrease in nuclear factor kappa-B (NF-kB) was recorded in piperine treated group. These findings indicate that administration of piperine ameliorated most of the tested biochemical changes induced by I/R in irradiated rat brain and serum.