7,8-Dihydroxyflavone (7,8-DHF), a member of the flavonoid family, has received considerable attention as a selective tyrosine kinase receptor B agonist. However, the pharmacological mechanisms responsible for its anti-inflammatory activities in microglial cells have yet to be elucidated. In this study, we evaluated the anti-inflammatory effects of this compound on the production of inflammatory mediators and cytokines in lipopolysaccharide (LPS)-stimulated murine BV2 microglial cells. At non-toxic concentrations, 7,8-DHF attenuated the production of nitric oxide (NO) and prostaglandin E2 (PGE2), by inhibiting inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) expression, respectively. Furthermore, the release and expression of inflammatory cytokines, including tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), were inhibited by 7,8-DHF. In addition, 7,8-DHF suppressed nuclear factor-κB (NF-κB) translocation and its transcriptional activity by blocking IκB (IκB)-α degradation; in addition, it exerted suppressive effects on the phosphorylation of mitogen-activated protein kinases (MAPKs). These results indicate that 7,8-DHF possesses therapeutic potential against neurodegenerative diseases that involve microglial activation.