Osteopotin Regulates The Expression Of MMP13 Through NF-?B Pathway In Osteoarthritis Synoviocytes

Objective:Osteopontin(OPN)is closely related to the severity and progression of Osteoarthritis(OA),but the role of OPN in the pathogenesis of OA is not clear.The purpose of this study was to investigate the role of osteopontin in knee osteoarthritis and its signal transduction pathway.Methods:15 patients with OA were selected,including 7 male and 8 female with an average age of 61.2�8.5 years.15 cases of control group were selected,including 9 male and 6 female with an average age of 42.4.�7.8 years with meniscus injury or ligament injury without osteoarthritis.OA synoviocyte was separated from digesting OA synoviocyte tissue,and taken first generation for training.According to the reported siRNA sequence,we synthetize OPN-siRNA as experimental group with OPN.Con-siRNA group was negative control group.Using Immunofluorescence detected the expression of intemal and extemal nuclear factor kappaB(NF-?B).Using Western Blotting detected the protein expression of NF-?B p65,p-NF-?B p65.Using IPP software deals with the Western blot images and reach the ash density value.We performed homogenization treatment of the target protein NF-?B/p-NF-?B and ?-actin.The results of NF-?B/p-NF-?B p65 were divided by the results of ?-actin,and the relative quantitative values of each group were obtained.OA synovial cells transfected with Con-siRNA were used as control group.OPN-siRNA,OPN,PDTC and PDTC combined with OPN were used as experimental group.Using Western Blotting detected the protein expression of MMP13.Result:Compared with control group,OPN treatment significantly increased the abundance of p65 and phosphorylated p65(P=0.0025,P=0.0019).The abundance of p65 and phosphorylated p65 in synoviocytes with OPN-siRNA treatment was significantly lower compared to the controls(P=0.0024,P=0.0176).PDTC treatment also inhibited translocation of p65 protein from the cytoplasm to the nucleus.OPN could significantly increase the protein abundance of MMP13(P=0.0075).OPN-siRNA can significantly reduce the protein abundance(P=0.0021).PDTC could significantly inhibit the regulation of OPN on MMP13.Conclusions:OPN can accelerate the transfer of NF-?B p65 from the cytoplasm to the nucleus in OA synovial cells,promote the expression of NF-?B p65,p-NF-?B p65,and promote the expression of MMP13 by activating the NF-?B pathway.