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 (5x105 (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.

Based on knowledge from traditional Iranian herbal medicine, this ex vitro study aims to examine the anti-inflammatory mechanism of Berberis vulgaris L by measuring the expression and release of pro-inflammatory cytokines, tumor necrosis factor-α (TNF-α), cyclooxygenase-1 (COX-1), and interleukine-18 (IL-6), IL-1β and inducible nitric oxide synthase (iNOS) in THP-1. The effects were assessed by measuring the levels of secretory proteins and mRNA of TNF-α and IL-6, the levels of nitric oxide (NO) secretion and the expression of iNOS in THP-1 cells. Cells were treated with 20 ng lipopolysaccharide/ml (LPS) in the presence and absence of concentrations of extracts from the Berberis vulgaris L. During the entire experimental period, we used extract concentrations (0.3 μg/mL) that had no cytotoxic effects, as measured with MTT, MTT and LDH assays. Berberis vulgaris extracts remarkably suppressed the LPS-induced NO release, significantly attenuated the LPS-induced transcription of iNOS and inhibited in a dose-dependent manner the expression and release of TNF-α. significant effects were observed on the release of IL-6. Taken together, these results suggest that Berberis vulgaris probably exerts anti-inflammatory effects through the suppression of TNF-α and iNOS expressions.

Abstract:Osteoarthritis (arthritis) is biomechanical, biochemical and cellular phenomenon, and is not known as a degenerative disease. Arthritis is one of the common chronic diseases and the most important reason of physical disability in the world. According to its side effect such as peptic ulcers, gastrointestinal bleeding, liver toxicity and renal complications dueofprescribing current treatment contain corticosteroid and nonsteroidal, we decided to evaluate possible effect of anti-inflammatory Esential oil of Fraxinus excelsior (EOFE) on biomarkers involved in disease. EOFE were prepared of genetic resources center. Bovine articular cartilage derived from the metacarpophalangeal joints of 14-18-month-old animals (without any sign of inflammation and bleeding) sent to laboratory in sterile bags at 4ºC. Cells were cultured in appropriate condition and counted by hemocytometer, viability assessed by trypan blue. After LPS treatment, cytokine levels were assayed. Cells cultures again and were kept in 37C, 90% humidity in CO 2 incubator and after RNA extraction, RT-PCR and PCR done. Also by Real-time PCR, gene expression was evaluated. E.E.F.E level cause down regulation of COX-2, IL-1β, TNF-α in LPS-stimulated cells.