Aim: To evaluate the effects of Pleurotus ostreatus on the lipid profile and atherogenic indices in Hyperlipidemic rats. Study Design, Place and Duration of Study: This case-control study was done for 60 days between March and April, 2017 at the department of Medical Laboratory Science and Department of Chemical Pathology, Babcock University, Ogun State, Nigeria. Methodology: Thirty male wistar rats weighing 117-130 g were divided randomly into 3 groups: Normolipidemic (NL) rats (fed with standard rodent chow), Hyperlipidemic (HL) rats (fed with standard rodent chow + duck yolk and reused oil), Hyperlipidemic Treated (HL+T) rats (fed with standard rodent chow + duck yolk and reused oil + 5% Pleurotus ostreatus powder).  Changes in the animal body weights were measured in this study. Serum was obtained from fasting blood samples for the standard biochemical analyses of total cholesterol (TC), triglycerides (TG), High density lipoprotein cholesterol (HDL-C), creatinine, urea, Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST). Low density lipoprotein (LDL), very low density lipoprotein (VLDL), TC/HDL, LDL/HDL and Log (TG/HDL) ratios were calculated. Results: The HL+T rats compared to HL rats had significantly reduced body weight, TC, TG, LDL, VLDL, TC/HDL, LDL/HDL and Log(TG/HDL) by 19.59%, 14.38%, 15.82%, 25.52%, 15.83%, 28.89%, 20.24% and 27.27% respectively (p ≤ 0.05) but recorded no significant change in HDL-C (p > 0.05). Creatinine, urea, AST and ALT did not show any significant change in HL rats and HL+T rats (p > 0.05). Conclusion: Treatment of hyperlipidemic male wistar rats with Pleurotus ostreatus reduced body weight, lipid levels (TC, TG, LDL, VLDL) and atherogenic indices (TC/HDL, LDL/HDL, Log (TG/HDL)) and appeared to have no detrimental effects on the liver and kidneys. These findings may provide insights and scientific basis for the promotion of the use of Pleurotus ostreatus in controlling hyperlipidemia and associated complications.