Effects of Icariin on IL-1β-induced rheumatoid arthritis synovial fibroblasts proliferation and TNF-α and RANKL production

Objective To observe the effects of Icariin(ICA) on proliferation of rheumatoid arthritis synovial fibroblasts(RASFs) stimulated by IL-1β and the expression of TNF-α and RANKL.Methods RASFs were isolated form Synovial tissue of RA patients and cultured in vitro;second-generation RASFs were plated in 96 / 6-well,setting with normal group、control group、MTX group、high-dose ICA group and low-dose ICA group;all groups except the normal,after stimulated by 5 ng/ml IL-1β for 24 hours,were continuously treated with different medicine for 72 hours.Then the RASFs proliferation was evaluated with WST-1 Cell Proliferation and Cytotoxicity Assay Kit,the expression of TNF-α and RANKL in supernatant of each group were detected by ELISA.Results Compared with normal group,Control group has a significantly higher absorbance values at 17.9% proliferation ratio(P0.05),also TNF-α and RANKL were higher expressed at the level of(361.542±7.178)ng/L and(428.468±11.118)ng/L respectively(P0.05).Compared with control group after being intervened for 72 hours,MTX can inhibit the proliferation of RASFs(P0.05),and reduce the production of TNF-α and RANKL while without significant difference(P0.05);RANKL levels in IH group was(375.766±7.806) ng/L(P0.05);low-dose ICA can inhibit RASFs proliferation with a proliferation ratio of-31.90%,and down-regulate the concentration of TNF-α at(334.542±6.691)ng/L or RANKL(365.495±5.280)ng/L(P0.05).Conclusion IL-1β can promote proliferation of RASFs and stimulate the production of cytokines through TNF-α and RANKL;icariin has the effect in inhibiting proliferation of IL-1β-stimulating RASFs and TNF-α,RANKL production.