Introduction: Global warming is one of the major issues in the current scenario. Heat stress is known to affect both physical and mental task. Imbalance in the maintenance of temperature leads to oxidative stress and damage to the body systems. Several studies have been documented on various stressors and its impact on antioxidant status. Aim: The present study was done to evaluate the antioxidant status in rats after acute heat stress. Methodology: Twelve male Swiss albino rats were randomly divided into two groups, control group, and experimental group. The experimental group of rats was exposed to acute heat stress at 38 ºC for four hours for a single day. Blood from the jugular vein was collected for the estimation of antioxidant statuses like LPO and SOD, GPx, CAT, vitamin C, and vitamin E in rats. Results: After exposure to acute heat stress, there was a significant and progressive increase (P<0.05) in LPO levels of the experimental group than compared to controls. Superoxide dismutase (SOD), Glutathione peroxidase (GPx) Catalase, vitamin C, and vitamin E showed a significant decrease (P<0.05) in the experimental group compared to the control group. The findings of the present study suggest that oxidative stress increases progressively, and the structural damage to the tissues may be due to oxidative stress.