The Study On Anti-osteoarthritic Effects Of Costunolide In Vitro And In Vivo

Objectives:The research to explore the therapeutic effect of costunolide on osteoarthritis(OA)and its intrinsic mechanism through in vivo and in vitro experiments.Methods:In the in vitro experiments,the rat knee joint chondrocytes were extracted for study.Interleukin-1?(IL-1?)was used to stimulate the chondrocytes to establish a cell model of osteoarthritis,and medial meniscus were taken off from the knees of rats to create the animal model of osteoarthritis.real-time polymerase chain reaction(q RT-PCR)was used to investigate the m RNA expression of COX-2,INOS,MMP-3,MMP-9 and MMP-13 of chondrocytes induced by IL-1? at different concentrations of costunolide.Western Blot experiments were used to detect the protein expressions of COX-2,INOS,IL-6,MMP-3,MMP-9,and MMP-13 in chondrocytes induced by IL-1?under the impact of different concentrations of costunolide.Western Blot experiments and immunofluorescence experiments were conducted to investigate the effects of costunolide on IL-1?-activated NF-?B and Wnt /?-catenin signaling pathways.The vivoexperiments showed differences between the costunolide-treated group and the OA group by Mankin score and immunohistochemistry.Results:In vitro experiments,costunolide had no significant effect on chondrocyte viability and phenotypic maintenance at a concentration of 0-6 ?M.And costunolide reduced IL-1?-induced the expression of MMP3,MMP9,MMP13,COX-2,IL-6 and INOS.costunolide can significantly reduce p65 phosphorylation and the location of p65 into nucleus in rat chondrocytes induced by IL-1?,thereby inhibiting the activation of NF-?B signaling pathway.At the same time,the activation of ?-catenin induced by IL-1? can also be inhibited by costunolide,thereby preventing its activated form being transferred into the nucleus to activate the expression of target genes.In vivo experiments,immunohistochemical methods showed that the costunolide group had lower MMP-13 and COX-2 expression in chondrocytes than the OA group,as well as the costunolide group showed less seriously cartilage degeneration and lower Mankin score.Conclusion:In our research,we confirmed that costunolide can exert anti-inflammatory effects and inhibit MMPs expression through the wnt / ?-catenin and NF-?B signaling pathways,thereby inhibiting the degradation of cartilage matrix and delaying the development of osteoarthritis.Taken together,these results indicate that costunolide have potential therapeutic value in the treatment of OA.