Anti-inflammatory effects of Clematis chinensis Osbeck extract(AR-6) may be associated with NF-κB, TNF-α, and COX-2 in collagen-induced arthritis in rat

The root of Clematis chinensis Osbeck has been used widely in rheumatoid arthritis in Chinese traditional medicine, and AR-6 is a triterpene saponin isolated from it. In this present study, we investigated the in vivo effects of oral AR-6 in chronic rat with collagen-induced arthritis (CIA) and possible molecular mechanism. CIA was induced by immunizing 56 female Sprague-Dawley (SD) rats with chicken typeIIcollagen (CII). Following eighteen days, the immunization rats with CIA were treated with AR-6 (32, 16, 8 mg/kg), cyclophosphamide (7 mg/kg), and TGP (Total Glucosides of Paeonia) (180 mg/kg) for 7 days, and rats without CIA were given the same volume of purified water. TNF-α and IL-1β levels in peripheral blood will be measured by ELISA, and Western blot analysis will be used to detect the expression of NF-κB p65 subunits, TNF-α and COX-2, in synovial membrane. We found that therapeutic treatment with AR-6 markedly improves the paw swelling and histopathological changes. Moreover, the serum levels of pro-inflammatory cytokines TNF-α and IL-1β were markedly lowered, and the expression of NF-κB p65 subunits, TNF-α and COX-2, in the synovial membrane of CIA rats was significantly inhibited in the AR-6-treated groups. These results enable to prove that AR-6 has a potential anti-inflammatory effect in CIA rats, and its mechanism may relate to the inhibition of the expression of NF-κB p65 subunits, TNF-α and COX-2.