In the present study, an anthraquinone derivative, questinol was successfully isolated from the broth extract of the marine-derived fungus Eurotium amstelodami for the first time. The structure of questinol was determined based on the analysis of the MS and NMR spectral data as well as comparison of those data with the published data. Moreover, the anti-inflammatory effect of questinol in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells was investigated. The results showed that questinol did not exhibit cytotoxicity in LPS-stimulated RAW 264.7 cells up to $200{mu}M$. Questinol could significantly inhibit NO and $PGE_2$ production at indicated concentrations. Questinol was also found to inhibit the production of pro-inflammatory cytokines, including TNF-${alpha}$, IL-1${eta}$, and IL-6. Furthermore, the western blot analysis showed that questinol suppressed the expression level of iNOS in a dose-dependent manner. However, questinol could slightly inhibit the expression of COX-2 at the concentration of $200{mu}M$. Therefore, our study suggests that questinol might be selected as a promising agent for the prevention and therapy of inflammatory disease.