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Azadirachta indica A. Juss. (Mimba) has been widely used as a traditional medicine to treat various diseases in Indonesia. However, its underlying mechanism as an anti-inflammatory agent is still unknown. The aim of this study was to identify the anti-inflammatory activity of ethanolic extract of Azadirachta indica A. Juss. leaves (AIL) and to assess its possible mechanism. The research initiated by extracting the active contents of AIL by maceration using ethanol 96 %. Cytotoxicity was observed by using MTT assay on RAW 264.7 cells. The effect of AIL on macrophages was determined by measuring NO production levels in the culture supernatant by using Griess assay. The results showed that AIL possesses a cytotoxic effect on RAW 264.7 cells with IC50 value of 195,01 µg/ml. The results also demonstrated that NO production levels were significantly decreased after AIL treatment. These findings suggested that AIL might stimulate anti-inflammatory activity. Further study are necessary to evaluate the anti-inflammatory activity of AIL deeply.