In this study, we described the isolation of an 8,14-secoergostane-type, a 9,11-secoergostane-type, and three ergostane-type steroids from the fruiting bodies of Pleurotus cornucopiae var. citrinopileatus. The structure of (22Z)-3β,5α,11-trihydroxy-9,11-secoergosta-7,22-diene-6,9-dione, previously reported, have been revised to (22E). Their structures were established using NMR, UV, IR, and mass spectroscopic analyses. Three of the isolated compounds were found to exhibit inhibitory activity on the production of nitric oxide in lipopolysaccharide-stimulated RAW264.7 macrophages with IC values of 21.3, 17.6, and 23.1 μM, respectively.