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Context: Pain-relieving plaster (PRP) is a traditional Chinese medicine (TCM) that has been widely used with satisfactory results in the treatment of some diseases related to inflammation, such as bruises, chronic arthritis. Objective: The mechanisms underlying the anti-inflammatory actions of PRP are investigated in this study for the first time. Materials and methods: The anti-inflammatory effects of PRP extracts were evaluated in lipopolysaccharide (LPS) or calcium ionophore A23187-treated murine peritoneal macrophages (PMs). Tumor necrosis factor-αÎł (TNF-αÎł), interleukin-1βÎø (IL-1βÎø), prostaglandin E<sub>2</sub> (PGE<sub>2</sub>), and leukotrienes B<sub>4</sub> (LTB<sub>4</sub>) were evaluated by ELISA assays. Reverse transcriptase-polymerase chain reaction (RT-PCR) and western blot analysis were used to detect the expression of cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX). Nuclear factor-kappa B (NF-κÎðB)--DNA-binding activity was determined by gel mobility shift assay. Results: PRP extracts were found to inhibit the production of TNF-αÎł, IL-1βÎø, and PGE<sub>2</sub>, reduce the expressions of COX-2 at the mRNA and protein levels induced by LPS, and reduced the production of LTB<sub>4</sub> induced by A23187. Furthermore, PRP extracts significantly attenuated LPS-induced NF-κÎðB--DNA-binding activity. Discussion and conclusion: The anti-inflammatory effects of PRP possibly are related to reduction of inflammatory cytokines (TNF-αÎł and IL-1βÎø), inducible inflammatory enzyme (COX-2), and its metabolite PGE<sub>2</sub> via NF-κÎðB signal pathway. Moreover, PRP extracts also notably inhibited the production of LTB<sub>4</sub>, indicating that PRP inhibited the 5-LOX pathway, which may be the other mechanism for its anti-inflammatory action.