| Background Osteoarthritis（OA）is a chronic,low-grade inflammatory disease that is based on the degeneration of cartilage and accompanied by sclerosis of subchondral bone,perichondrium hyperplasia and ossification,synovitis,ligament injury around the joint and other changes in joint structure.The early clinical manifestations are intermittent pain related to movement,and the late clinical manifestations are persistent pain and significant movement limitation or even deformity.At present,it is conservatively estimated that there are about 100 million OA patients in China,and with the change of China’s age structure and aging population,the number of OA patients will continue to increase in the future.At present,clinical treatment is mainly aimed at pain,and there is no effective treatment.Therefore,it has been a hotspot of orthopaedic research to explore the pathophysiological mechanism of OA and find effective treatment methods.Conventional wisdom is that OA is a kind of diseases of "mechanical wear,the current study found that cartilage catabolism and anabolism balance disorder is one of the important factors in cartilage degeneration,therefore explore the regulation of catabolism and anabolism pathway,restore the balance of catabolism and anabolism is looking for a new scheme is an effective means of OA treatment.Daphnetin is a kind of benzopyranone compound extracted from Chinese traditional medicine Cangbai ruixiang,and is the main effective component of Zushima injection.Clinical studies have proved that Zushima injection can effectively treat arthritis,but its mechanism of action is still unclear.Therefore,the application value of daphnetin in the treatment of arthritis deserves further study.ObjectiveBased on the regulation of anabolism and catabolism of articular cartilage,to explore whether DAP can delay the progression of OA and its mechanism.Methods（1）From the joint replacement abandoned cartilage primitive chondrocytes,cultivate people with toluidine blue staining method,identified by immunofluorescence cell,identified as cartilage cell,with CCK 8（cell counting kit 8）of different concentration Daphnetin（Daphnetin,DAP）on the viability of chondrocytes.（2）In vitro OA model was constructed with IL-1β.RNA and protein were extracted after stimulation with different concentrations of daphnetin.The extracted RNA was used for fluorescence quantitative PCR（q RT-PCR）experiment.The extracted proteins（nucleoprotein and plasma protein）were used for Western blot assay.In this study,the catabolic indexes of chondrocytes treated by DAP were detected: MMP-1,MMP-13 and ADAMTS-5.Anabolic indicators: whether there were differences in the expressions of collagen-II,Aggrecan and OA chondrocytes,and the changes of NF-κB and MAPK signaling pathways.（3）OA in vivo model was made by using medial meniscus instability（DMM）,and after injection of daphne into the joint,Joint samples were extracted for Safranin O/Fast Green staining（Safranin O/Fast Green）test to determine whether there was a difference in cartilage degeneration between daphnetin-injected mice and mice with common arthritis.OARSI（Osteoarthritis Research Society International）score was used for semi-quantitative analysis to verify the therapeutic effect of daphnetin on OA.Results（1）DAP can decrease the expression levels of MMP-1,MMP-13 and ADAMTS-5 in IL-1β-induced OA model cells in vitro,and inhibit the catabolism of OA model cells in vitro.At the same time,the expression levels of collagen-II and Aggrecan were increased,which promoted the anabolism of OA model cells in vitro.（2）DAP inhibits Il-1 β-induced overactivation of NF-κB and MAPK signaling pathways in OA model cells in vitro.（3）DAP can delay the degeneration of cartilage in DDM mice OA.ConclusionDAP inhibits catabolism and promotes anabolism in human osteoarthritis chondrocytes,and alleviates knee cartilage degeneration,which may be related to its inhibition of NF-κB and MAPK signaling pathways. |
PDF Full Text Request