We investigated the effect of Orostachys japonicus extracts on serum lipids, leptin and insulin level in hyperlipidemic rats. Also, diacylglycerol acyltransferase (DGAT) activity and thiobarbituric acid reactive substance (TBARS) were assessed. Inhibitory effect of DGAT related to triglyceride synthesis emerged approximately 96% in EtOAc fraction and showed 90% and 67%, respectively, in CHCl3 and BuOH fractions. Furthermore, the EtOAc and BuOH fractions inhibited 81% and 77%, respectively, in glycerol-3-phosphate acyl transferase (GPAT). Hyperlipidemia and obesity marker, contents of leptin and insulin on serum of hyperlipidemic rats, decreased 50% and 25%, respectively, compared with control group in treated EtOAc fraction. The oxidative stress marker, a concentration of TBARS, showed decrease of approximately 30% in treated EtOAc fraction. Moreover, high density lipoprotein (HDL)-cholesterol contents on serum of rats fed a hyperlipidemic diet were increased 10% and low density lipoprotein (LDL)-cholesterol decreased 50% as well as triglyceride amount of feces multiplied approximately two times more than control group in treated EtOAc fraction. The data suggest that the fractions of O. japonicus may be a potent biomaterial for treatment of hyperlipidemia or obesity.