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The anti-inflammatory activity of ethanol extract from Chondrus nipponicus Yendo (CNYEE) was investigated by measuring production of a lipopolysaccharide-induced inflammatory response mediator. CNYEE had no cytotoxic effects on proliferation of macrophages compared to the control. CNYEE significantly inhibited (over 50%) NO production at 50 μg/mL, with inhibitory effects on expression levels of cytokines such as interleukin (IL)-6, tumor necrosis factor-α (TNF-α), and IL-1β. In particular, IL-6 inhibitory activity of CNYEE was higher than 70% at 100 μg/mL. CNYEE also reduced protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and nuclear factor (NF)-κB in a dose-dependent manner. CNYEE also significantly reduced phosphorylation of p38, extracellular signal-regulated kinase, and c-Jun N-terminal kinase. Therefore, these results suggest that CNYEE may have anti-inflammatory effects by modulating the NF-κB and mitogen-activated protein kinases signaling pathways and may be used as an anti-inflammatory therapeutic material.