For centuries, humans have leveraged the health-promoting properties of plants for our well-being. While research has been conducted on numerous medicinal plants, the specific benefits of many species remain underexplored. Eupatorium Japonicum (EJ), a member of the Asteraceae family, has historically been consumed in Japan, South Korea, China, and Vietnam for its traditional use in soothing digestive issues. This study aimed to explore the radical scavenging and antiinflammatory efficacy of EJ extract using RAW 264.7 cells. The radical-scavenging effects were assessed using the DPPH and ABTS assays, where an anti-oxidative molecule in the test sample will react with a stable free radical in DPPH and ABTS causing discoloration. The anti-inflammatory efficacy was assessed using the nitric oxide (NO) assay in LPS-induced RAW 264.7 cells, where the amount of NO produced in response to infection was measured using Griess reagent. Reversetranscriptase polymerase chain reaction (RT-PCR) and real-time PCR were executed to confirm the anti-inflammatory activity by measuring the RNA levels of pro-inflammatory cytokines. The DPPH and ABTS assays revealed that EJ extract decreased oxidation in a concentration-dependent manner (7.8-1,000 μg/mL) compared to ascorbic acid and Trolox respectively. EJ extract significantly reduced NO production concentration independently. Furthermore, EJ extract showed no cytotoxic effects as determined through the MTT assay. RT-PCR and real-time PCR analyses revealed inhibition of mRNA expression of pro-inflammatory cytokines (iNOS, COX-2, TNF-α, and IL-6). Western blotting demonstrated EJ’s anti-inflammatory activity by reducing protein levels of iNOS, COX-2, TNF-α, and IL-6. These findings suggest that EJ extract exhibits anti-inflammatory activities and can be further evaluated in the future.