PMPs通过CXCR2/Erk/NF-κB通路影响RA-FLS中RANKL和OPG表达的研究

PMPs regulate the expressions of RANKL and OPG in RA-FLS via CXCR2/Erk/NF-κB pathway

用不同浓度血小板微颗粒(PMPs,0、25、50μg·mL^(-1))分别处理RA-FLS,根据高通量基因芯片分析基因表达谱的改变,应用类风湿关节炎-成维样滑膜细胞(RT-qPCR)、Western blot和ELISA方法检测核因子κB受体活化因子(RANKL)和骨保护素(OPG)的表达;利用Western blot检测相关信号通路抑制剂对PMPs作用下类风湿关节炎-成纤维样滑膜细胞(RA-FLS)中核因子κB受体活化因子配体(RANKL)和OPG表达的影响。结果表明:PMPs可上调RA-FLS中RANKL表达,下调OPG表达;抑制NF-κB、CXCR2的活化能拮抗PMPs对RANKL和OPG表达的影响;PMPs上调RA-FLS中Erk的磷酸化水平时,IκB和NF-κB的磷酸化水平也随之上调。说明PMPs可能通过激活CXCR2/Erk/NF-κB信号通路上调RA-FLS中RANKL、下调OPG,参与类风湿关节炎的骨质破坏与重塑。

RA-FLS was treated with 0,25,50μg·mL^(-1)PMPs.According to the gene expression profile detected by high throughput gene chip,the expression of RANKL and OPG was examined by RT-qPCR,Western blot and ELISA assays.Combined with signal pathway inhibitors,the effect of PMPs on expressions of RANKL and OPG in RA-FLS wsd evaluated by Western blot.The results showed that PMPs up-regulated the expression of RANKL and down-regulated the expression of OPG in RA-FLS.The results also demonstrated that the inhibition on NF-κB or CXCR2 activation antagonized the effect of PMPs on expressions of RANKL and OPG,and when PMPs induced phosphorylation of Erk in RA-FLS,phosphorylations of IκB and NF-κB were induced,too.Data from this study indicated that PMPs can up-regulate the expression of RANKL and down-regulate the expression of OPG in RA-FLS.The mechanism may be related to activating CXCR2/Erk/NF-κB signaling pathway,which may provide a new idea for the treatment of rheumatoid arthritis.