Due to the side effects of current therapies for osteoarthritis one of the alternative medicine is using herbal medicine such as Nigella sativa .L which in Iranian traditional medicine has been used as a treatment option. The purpose of this study is evaluating the effect of alcoholic extract of Nigella sativa (AENS) on pro-inflammatory cytokines in Bovine Fibroblast-like BFLs (BFLS) and THP-1. BFLS cells were isolated from Radiocarpal joint. After evaluating of LC50 (27 µg/mL), both cells

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... 5 (cells\wel)) were incubated at 37 ° C and 5% CO2 and 90% humidity for 72 hours with AENS (6.13 μg\ml as a media LC50). One set of cells was activated for 1h with LPS for RT-PCR analysis of COX-2, INOS, IL-1β, TNF-α expression and another set of cells was activated for 24h, cells supernatant were analyzed for PGE-2 and nitrite content. The present study demonstrates that AENS reduced expression levels of COX-2, INOS, TNF-α in control group. Reduced expression of COX2 and INOS was significantly along with the reducing production of NO and PGE2.Also, AENS decreased the expression of TNF-α and iL-1β in control group. Our results showed that the anti-inflammatory effect of AENS not only has anti-inflammatory effect on the BFLS cells but also related to the THP-1 that are active in the synovial membrane.