Effect Of NF-?B Signaling Pathway In Osteoclastogenesis In Patients With Rheumatoid Arthritis After Exposed To Curcumin

Objective To observe the effect of Curcumin(Cur)on osteoclastogenesis in rheumatoid arthritis(RA)patients,and to explore the regulation mechanism of nuclear factor kappa B(NF-?B)signaling pathway during this process,to provide a theoretical reference for the prevention and treatment of bone destruction in RA.Methods(1)Peripheral blood mononuclear cells(PBMCs),separated from RA patients in vitro,were cultured into osteoclasts,and given different concentrations of Curcumin during the culture.(2)Groups:Control group,Cur 2.5 ?M group,Cur 5 ?M group and Cur 10 ?M group,given different concentrations of Curcumin.(3)After cultured for 14 days,tartrate-resistant acid phosphatase(TRAP)staining was used to mark the mature osteoclasts and counted the number of the mature osteoclasts in different groups.(4)Western blotting was used to detect the expression of I?B? and p-I?B? in cells in different groups.(5)Western blotting was used to detect the expression of NF-?B p65 in the cytoplasm and nucleus in different groups.(6)Immunofluorescence staining was used to detect the nuclear translocation of NF-?B p65.Results(1)After cultured for 14 days,each group of cells were consistent with the morphological characteristics of osteoclasts.(2)TRAP staining showed that the number of TRAP positive cells in Cur 2.5?M group,Cur 5 ?M group and Cur 10 ?M group was significantly lower than that in control group,and the difference was statistically significant(P<0.05).(3)Western Blotting showed that the expression of I?B? in Cur 2.5?M group,Cur 5 ?M group and Cur 10 ?M group was significantly higher than that in Control group,and the expression of p-I?B? was significantly decreased(P<0.05).(4)Compared with the control group,the expression level of NF-?B p65 in cytoplasm in Cur 2.5 ?M group,Cur 5 ?M group and Cur 10?M group was significantly increased,and the expression level of NF-?B p65 in nucleoprotein was significantly decreased(P<0.05).(5)Immunofluorescence staining showed that NF-?B p65 was located predominantly in the cytoplasm in an unstimulated state.Stimulation with RANKL caused nuclear translocation of p65.Pretreatment with Curcumin at a concentration of 5 ?M weakened the nuclear accumulation of p65.Conclusion Curcumin inhibited the osteoclastogenesis in RA patients by inhibiting of NF-?B activation,which was accompanied with the phosphorylation and degradation of I?B?and the decrease of nuclear translocation of NF-?B p65.