Influence of CXCL13 on expression of RANKL in human synovial cells induced by TNF-α

JIN Shengyu; REN Hongge

doi:10.3969/j.issn.1002-266X.2017.48.002

Abstract：Objective To investigate the effect of CXCL13 on the expression of receptor activator of NF-κB ligand (RANKL) in human synovial cells induced by tumor necrosis factor-α (TNF-α).Methods ① The human synovial cells were cultured in vitro and were treated with 10 g/mL TNF-α.At 0,6,12,and 24 h,we used immunofluorescence to detect the CXCL13 expression.② The human synovial cells cultured in vitro were treated with 10 g/mL TNF-α for 24 h,after that,we replaced the culture fluid with medium containing 0,5,10,and 25 ng/mL CXCL13 and continued to culture for 24 h,or replaced the culture fluid with medium containing 25 ng/mL CXCL13 and continued to culture for 0,1,3,6,and 24 h.The relative expression of RANKL was detected by Western blotting.The normal cultured cells without treatment of TNF-α and CXCL13 were used as the control group.Results ①The expression levels of CXCL1 3 (fluorescence intensity) were 0.907 ± 351,0.823 ± 735,0.710 ± 664,and 0.653 ± 853 in cells treated with TNF-α at 0,6,12,and 24 h (all P ＜ 0.05).②The expression of RANKL protein was 0.956 ± 0.014 in the control group,and the expression levels of RANKL protein in cells treated with 25 ng/mL CXCL13 for 0,1,3,6,and 24 h were 1.543 ± 0.047,1.366 ± 0.026,0.883 ± 0.026,0.367 ± 0.034,and 0.246 ± 0.015;the expression levels of RANKL protein in cells treated with different contentrations (0,5,10,25 ng/mL) of CXCL13 were 0.287 ±0.007,0.189 ±0.008,0.069 ±0.004,and 0.022 ± 0.002;significant difference was found between them (all P ＜ 0.05).Conclusion CXCL13 can inhibit the expression of RANKL protein of osteoarticular synovial cells induced by TNF-α in a certain concentration and time.