The effects of ergosterol-rich Lentinus edodes stalk extracts on lipid metabolism remain unknown; thus, the content of ergosterol in the cap and stalk of L. edodes was analyzed using high performance liquid chromatography and the regulatory effects of the n-hexane extract (SEHE) and methanol/dichloromethane extract (SEMD) of the stalk on serum lipids, liver lipids and fecal lipids investigated in a rat model induced by a high-cholesterol diet. The ergosterol content of 7.82 mg∙g-1 in the L. edodes cap was found to be significantly higher than the 5.24 mg∙g-1 in the stalk, and the ergosterol content of SEHE was 20.31% (m/m) was higher than the 6.12% (m/m) obtained for SEMD. Animal experiments showed that both SEHE and SEMD could significantly reduce the level of low-density lipoprotein cholesterol (LDL-C), with reductions of 51.02% (P<0.05) and 58.34% (P<0.05), respectively, and effectively reduce the LDL-C/high-density lipoprotein cholesterol (HDL-C) ratio, with reductions of 54.61% (P<0.05) and 53.04% (P<0.05), respectively, in the rat serum. The two extracts were also found to reduce the cholesterol content (-19.31%, P<0.05; -23.73%, P<0.05) and triglyceride content (-9.82%, P=0.11; -24.83%, P<0.05) in rat liver. SEMD was also found to significantly reduce the total cholesterol (TC) level in rat serum (-13.31%, P<0.05). These findings indicate that both stalk extracts effectively regulated lipid metabolism in rats and have the potential to be used as functional food ingredients for the prevention of cardiovascular disease.