1,25二羟基维生素D3对强直性脊柱炎巨噬细胞极化的作用

Role of 1,25-dihydroxyvitamin D3 on the polarization of macrophages in ankylosing spondylitis

[目的]探讨1,25二羟基维生素D3对强直性脊柱炎(ankylosing spondylitis, AS)患者巨噬细胞极化的作用以及其对Toll样受体4 (toll-like receptor 4, TLR4)/髓样分化因子88 (myeloid differentiation factor 88, MyD88)/核因子-κB (nuclear factor-κB, NF-κB)信号通路的调节。[方法]以佛波酯诱导人单核细胞株THP1分化为巨噬细胞。正常组加入5%的健康体检者的血清,AS组加入5%的AS患者的血清,试验组加入5%的AS患者的血清,再添加1 ml的100 nmol/L的1,25-二羟基维生素D3溶液。ELISA法检测各组细胞上清液中IL-10、IL-6、TNF-α的含量。Western blot检测各组细胞信号通路相关蛋白的表达。[结果]与正常组相比,AS组和试验组细胞上清液中IL-10的含量明显下降(P<0.05),细胞上清液中IL-6、TNF-α的含量,细胞中TLR4、MyD88、NF-κB的表达明显增加(P<0.05);与AS组比较,试验组细胞上清液中IL-10的含量明显增加(P<0.05),细胞上清液中IL-6、TNF-α、细胞中TLR4、MyD88、NF-κB的表达的含量明显降低(P<0.05)。[结论] 1,25-二羟基维生素D3可能通过TLR4/MyD88/NF-κB信号通路,调节强直性脊柱炎患者巨噬细胞的极化过程。

[Objective] To investigate the effect of 1,25 dihydroxyvitamin D3 on macrophage polarization in ankylosing spondylitis(AS)and its regulation mechanism by Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB) signal pathway. [Methods] Phorbol ester was used to induce human monocyte line THP1 differentiate into macrophages. Subsequently, the cells were cultured with 5% serum of healthy subjects(normal group), 5% AS patient’s serum(the AS group), whereas 5% serum of AS patients,and 1 ml of 100 nmol/L 1,25-dihydroxyvitamin D3 solution(the trial group). ELISA method was used to detect the content of interleukin-10(IL-10), IL-6 and tumor necrosis factor-α(TNF-α) in the supernatant of each group of cells. Western blot was used to detect the cell signaling pathway related protein expression. [Results] Compared with the normal group, the AS group and the trial group had significantly decreased IL-10 in the cell supernatant, while significantly increased IL-6 and TNF-α in the cell supernatant, as well as the expression of TLR4, MyD88, NF-κB, which were statistically significant(all P<0.05). Compared with the AS group, the trial group had significantly increased IL-10 content in the cell supernatant, whereas significantly decreased IL-6 and TNF-α, as well as expression levels of TLR4,MyD88, and NF-κB with statistically significant differences(P<0.05). [Conclusion] 1,25-dihydroxyvitamin D3 might regulate polarization process of macrophages in ankylosing spondylitis by the TLR4/MyD88/NF-κB signaling pathway.