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Background/Objectives: This study aims to demonstrate pro-inflammatory cytokine production inhibitory effects using broccoli extract. Methods/Statistical analysis: Thus, This paper was carried out to see the production of NO and pro-inflammatory cytokine ,TNF-α and IL-6 using the macrophages of LPS-treated mice. The cytotoxicity test was used 96 AQueous One solution cell proliferation assay. NO was measured using NO Detection Kit, and the production of pro-inflammatory cytokine was measured using ELISA kit. Findings: As a result, broccoli extract had no cytotoxicity at 10ug/mL to 1,000 ug/mL and significantly inhibited the production of NO as well as TNF-α and IL-6 which are pro-inflammatory cytokines. In the RAW 264.7 macrpphage cells, Pro-inflammatory cytokine production inhibitory effects are likely to be diversely utilized as basic physiological activity data and functional materials to demonstrate anti-inflammatory properties of broccoli extract.Improvements/Applications: In conclusion, this study can be used to a basic data to objectively demonstrate the physiological activity of immunological mechanism associated with the anti-inflammatory action of broccoli extract. However, in-depth research on anti-inflammatory is needed.