The present study was designed to investigate the hypolipidemic effect of pumpkin seed oil in experimental hyperlipidemic rats. Fifty male rats were divided into five equal groups. Group 1: normal rats fed on normal diet. Group 2: (hyperlipidemic group) rats administered standard diet+ 20%coconut oil and 1% cholesterol daily for eight weeks. Group 3: (hyperlipidemic +pumpkin seed oil treated group) rats administered standard diet+ 20%coconut oil and 1%  cholesterol daily for four weeks followed by atherogenic diet+ 50 mg/Kg b. wt. pumpkin seed oil daily for other four weeks. Group 4: (hyperlipidemic +pumpkin seed oil protected group) rats administered standard diet+ 50 mg/Kg b. wt. pumpkin seed oil daily for two weeks followed by standard diet+ 20%coconut oil and 1%  cholesterol+ 50 mg/Kg b. wt. pumpkin seed oil daily for eight weeks. Group 5 : (pumpkin seed oil group) rats administered standard diet+ 50 mg/Kg b. wt. pumpkin seed oil daily for eight weeks. Blood samples collected for serum separation and used for determination of lipids profile (total cholesterol, triacylglycerol, LDL-C, HDL-C, VLDL-C, total lipids and phospholipids), liver marker enzymes (ALT and AST), albumin, kidney function markers (urea and creatinine) and glucose concentration. The results revealed that in group 2 there were significant increases in serum total cholesterol, triglycerides, LDL-C, VLDL-C, phospholipids, total lipids, ALT, AST, urea, creatinine and glucose while HDL-C and albumin were significantly decreased. Meanwhile, administration of pumpkin seed oil resulted in significant decrease in all elevated mentioned parameters and increase in HDL-C and albumin levels. Therefore, it could be concluded that pumpkin seed oil has hypolipidemic and hypoglycemic effect in rats fed high fat diet.