维生素D3对强直性脊柱炎患者血清诱导单核巨噬细胞极化的影响

目的: 研究维生素D3干预对强直性脊柱炎（AS）患者血清单核巨噬细胞极化的影响。方法: 选择2016年1月至2017年12月在温州市人民医院尚未治疗的AS初诊患者20例和健康志愿者20名。通过佛波酯诱导人单核-白血病细胞THP1分化为单核巨噬细胞，并与AS患者（AS对照组）或健康志愿者（健康对照组）的外周血血清联合培养；在含有AS外周血血清的混合培养基中加入维生素D3进行干预（维生素D3组）。流式细胞仪测定CD68和CD206+单核巨噬细胞的比例，RT-PCR测定精氨酸酶1（Arg-1）和诱导型一氧化氮合酶（iNOS）mRNA的表达水平。结果: 佛波酯可将THP1细胞诱导分化为单核巨噬细胞并表现出明显的细胞形态变化。AS对照组中CD206+细胞的比例低于健康对照组（t=9.434，P < 0.05），而CD68+细胞的比例高于健康对照组（t=43.920，P < 0.05）；维生素D3组中CD206+细胞的比例高于AS对照组（t=8.895，P < 0.05），而CD68+细胞的比例低于AS对照组（t=9.089，P < 0.05）。AS对照组中Arg1 mRNA表达量低于健康对照组（t=8.899，P < 0.05），而iNOS mRNA表达量高于健康对照组（t=3.656，P < 0.05）；维生素D3组Arg-1 mRNA表达量高于AS对照组（t=6.219，P < 0.05），而iNOS mRNA表达量低于AS对照组（t=5.876，P < 0.05）。结论: 维生素D3可能通过影响单核巨噬细胞的极化参与AS患者的免疫调节。
Abstract: Objective: To investigate the effect of vitamin D3 on polarization of monocyte macrophages induced by serum from patients with ankylosing spondylitis (AS). Methods: Twenty AS na？ve patients and 20 healthy controls from Wenzhou People's Hospital during January 2016 and December 2017 were enrolled. The macrophages were differentiated from THP1 cells induced by phorbol 12-myristate 13-acetate (PMA), and then co-cultured with the serum from healthy subjects (control group) or AS patients. Vitamin D3 was added in the medium mixed with serum from AS patients. Flow cytometry was used to analyze the ratio of CD68 and CD206 positive cells, and RT-PCR was performed to detect the mRNA expression of inducible nitric oxide synthase(iNOS) and arginase-1(Arg-1). Results: THP1 cells could be polarized into mononuclear-macrophages with the induction of PMA. The proportion of CD206 positive cells in AS-serum group was lower than that in the control group (t=9.434, P < 0.05), while the proportion of CD68 positive cells was higher than that in the control group (t=43.920, P < 0.05). The proportion of CD206 positive cells in vitamin D3 group was higher than that in AS-serum group (t=8.895, P < 0.05), while the proportion of CD68 positive cells was lower than that in AS-serum group (t=9.089, P < 0.05). mRNA expression of Arg-1 in AS-serum group was lower than that in the control group (t=8.899, P < 0.05), while mRNA expression of iNOS was higher than that in the control group (t=3.656, P < 0.05). mRNA expression of Arg-1 in vitamin D3 group was higher than that in AS-serum group (t=6.219, P < 0.05), while mRNA expression of iNOS was lower than that in AS-serum group (t=5.876, P < 0.05). Conclusion: Vitamin D3 can regulate the polarization of mononuclear macrophages for immunoregulation in patients with AS. Key words: Spondylitis,