青藤碱对类风湿关节炎患者外周血单核源性树突状细胞趋化因子分泌和受体表达的影响

Effect of sinomenine on the expression of chemokines and chemokine receptors in dendritic cells from patients with rheumatoid arthritis

目的观察中药单体青藤碱对类风湿关节炎患者外周血单核源性树突状细胞趋化因子分泌和受体表达的影响。方法分离8例类风湿关节炎患者的单核细胞,采用细胞因子刺激分化为树突状细胞。分别采用青藤碱高(5mmol/L)、中(2mmol/L)、低(1mmol/L)剂量和空白对照干预。流式细胞仪检测各组树突状细胞趋化因子受体CCR5和CCR7的表达,半定量RT-PCR检测树突状细胞CCR5和CCR7 mRNA的表达,酶联免疫吸附试验(ELISA)检测培养上清中趋化因子CXCL9(MIG)、CXCL10(IP-10)、CXCL11(ITAC)的表达。结果与对照组相比,经青藤碱高干预后的树突状细胞趋化因子受体CCR5、CCR7的蛋白和mRNA表达明显降低(P<0.05或P<0.01)。青藤碱干预后的DC分泌CXCL9(MIG)、CXCL10(IP-10)降低(P<0.05或P<0.01),但对CXCL11(ITAC)的表达没有影响。结论青藤碱可能通过抑制趋化因子对树突状细胞的趋化作用,减少其趋化因子的分泌,而产生治疗类风湿关节炎的作用。

Objective To investigate the effect of sinomenine on the expression of chemokines and chemokine receptors of dendritic cells in patients with rheumatoid arthritis in vitro. Methods The peripheral blood mononuclear cells isolated from 8 patients with rheumatoid arthritis were induced to differentiate into dendritic cells with GM-CSF and IL-4. The dendritic cells were exposed to sinomenine at high （5 mmol/L）, moderate （2 mmol/L）, and low （1 mmol/L） concentrations or treated with the control medium. The expression of CCR5 and CCR7 on the surface of the dendritic cells were measured by flow cytometry,. and the CCR5 and CCR7 mRNA expressions were detected by semi-quantitative PCR. Enzyme-linked immunosorbent assay （ELISA） was used to measure the expressions of CXCL9 （MIG）, CXCL10 （IP-10） and CXCL11 （ITAC）. Result Compared with the control cells, the dendritic cells treated with sinomenine, especially at high and moderate concentrations, showed significantly lowered mRNA and protein expressions of CCR5 and CCR7. Similar results were observed in the expressions of CXCL9 （MIG） and CXCL10 （IP-10）, but not in CXCL11 （ITAC）. Conclusion Sinomenine produces therapeutic effect on rheumatoid arthritis possibly by inhibiting the expression of chemokines and chemokine receptors in the dendritic cells to suppress the chemotactic migration of the dendritic cells.