Oxidative stress is a phenomenon caused by an imbalance between production and accumulation of oxygen reactive species (ROS) in the cells and tissues and the ability of a biological system to detoxify these reactive products. This study investigated the acute effect of different doses of ethanolic extract annabis sativa (EECS) on oxidative stress biomarkers in male (M) and female (F) Wister rats. Twenty male rats (170 g ± 1.24) and twenty female rats (150 g ± 1.05) were separately assigned into four groups of five animals each for male and female, such that the rats in groups IM and IF, IIM and IIF, IIIM and IIIF and IVM and IVF received orally 1 mL of distilled water, 2 mg/kg body weight (BW) of EECS, 4 mg/kg BW of EECS and 6 mg/kg BW of EECS respectively for twenty one (21) days. Catalase, superoxide dismutase (SOD), Glutathione peroxidase (GPx), Glutathione reductase (GSH), malondialdehyde (MDA), total antioxidant capacity (TAC) and lactate dehydrogenase (LDH) were determined using standard methods. Administration of different doses (2 mg/kg BW, 4 mg/kg BW and 6 mg/kg BW) of EECS significantly (p<0.05) decreased catalase, SOD, GPx, GSH and TAC levels respectively when compared with the control. However, Cannabis sativa (CS) increased MDA and LDH levels significantly (p<0.05) when compared with the control. It was deduced that these alterations in oxidative stress biomarkers were dependent on the doses of CS consumed. Thus, the more the concentration of CS consumed, the more it may affect oxidative stress biomarkers negatively. In addition, all these effects of CS on oxidative stress biomarkers were more in male than in female.