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

摘要:

[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 fac- tor-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。

Abstract:

[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 interleukin10 (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 in- creased 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.