Abstract: This study was undertaken to investigate the putative protective effect with antioxidant potential of the oyster mushroom (Pleurotus ostreatus) in glycerol-induced Acute Renal Failure (ARF) in rats. Four groups of rats were employed in this study, group I served as control, group II was given a single intramuscular dose of 50% glycerol in olive oil (8 mL kg-1 b.wt.), group III was given mushroom in the diet (62.5 g kg-1) and in drinking water (2.5 g L-1) for 3 weeks and group IV was given mushroom as the third group then was given a single intramuscular dose of 50% glycerol in the day 22 of mushroom administration. Oxidative stress markers in kidney, kidney abnormalities as well as hematological alteration were estimated. Glycerol treatment resulted in a marked renal oxidative stress, a significantly deranged renal function and a reduction in all hematological tested parameters. Pre-treatment with mushroom markedly reduced elevated thiobarbituric acid reacting substances, reduced glutathione, protein carbonyl and resorted the depleted renal antioxidant enzymes, attenuated renal dysfunction and the disturbances observed in hematological and biochemical parameters. These results suggest that Pleurotus ostreatus may have ability to protect the renal damage involved in acute renal failure in rats.