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<i>Weigela subsessilis</i> is used in folk medicine to treat pain and allergic syndromes in Korea. However, the antibacterial and anti-inflammatory activities of <i>W. subsessilis</i> callus extract remain unexplored. In this study, we aimed to evaluate the <i>W. subsessilis</i> callus of pharmacological activity. Therefore, we first established in vitro calluses of <i>W.</i><i>subsessilis</i> via plant tissue culture methods. We then evaluated the antioxidant and anti-inflammatory effects of <i>W. subsessilis</i> callus extract in lipopolysaccharide (LPS)-treated RAW264.7 macrophage cells. The <i>W. subsessilis</i> callus extract showed antioxidant and anti-inflammatory effects. These effects were regulated via suppression of mitogen-activated protein kinase signaling through LPS-induced translocation of nuclear factor kappa B (NF-κB) p65 from the cytoplasm to the nucleus. <i>W. subsessilis</i> callus extract also showed antibacterial and anti-inflammatory activities in <i>Propionibacterium acnes</i>-treated HaCaT keratinocyte cells. These results indicate that <i>W. subsessilis</i> callus extract has antioxidant, antibacterial and anti-inflammatory activities, suggesting its possible application in the treatment of inflammatory disorders.