Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus, Denmark.
Department of Endocrinology MEA, Aarhus University Hospital, Aarhus, Denmark.
PLoS One. 2019 Apr 12;14(4):e0215383. doi: 10.1371/journal.pone.0215383. eCollection 2019.
We wanted to investigate effects of vitamin D3 (25(OH)D3 and 1.25(OH)2D3) on inflammatory cytokine expression in both activated and non-activated Mφ.
Mononuclear cells, isolated from healthy donor buffy coats were cultured for a 6-day differentiation-period. Fully differentiated Mφ were pre-treated with either 25(OH)D3 or 1.25(OH)2D3 for (4, 12 or 24 hours) +/-LPS challenge for 4 hours. Gene expression analyses of VDR, Cyp27b1 and pro-inflammatory markers TNF-α, IL-6, NF-κB, MCP-1, was performed using RT-quantitative PCR. TNF-α protein levels from Mφ culture media were analysed by ELISA.
Both 25(OH)D3 and 1.25(OH)2D3 significantly inhibited TNF-α expression in both LPS-stimulated and unstimulated Mφ. Also, NF-κB, and to a lesser extend IL-6 and MCP-1 were inhibited. LPS up-regulated Cyp27b1 gene expression which was partly reverted by 1.25(OH)2D3.
These data show anti-inflammatory effects of vitamin D3 (25(OH)D3 and 1.25(OH)2D3) in human macrophages, and support, that means for targeting high dose vitamin D3 to the immune system may have beneficial clinical effect in inflammatory conditions.
我们旨在研究维生素 D3（25（OH）D3 和 1.25（OH）2D3）对激活和非激活巨噬细胞（Mφ）中炎症细胞因子表达的影响。
从健康供体的白细胞中分离出单核细胞，进行为期 6 天的分化培养。将完全分化的 Mφ 用 25（OH）D3 或 1.25（OH）2D3 预处理（4、12 或 24 小时），再用 LPS 刺激 4 小时。采用 RT-qPCR 检测 VDR、Cyp27b1 和促炎标志物 TNF-α、IL-6、NF-κB、MCP-1 的基因表达情况。通过 ELISA 分析 Mφ 培养物中 TNF-α 蛋白水平。
25（OH）D3 和 1.25（OH）2D3 均显著抑制 LPS 刺激和未刺激 Mφ 中的 TNF-α 表达。同时，NF-κB 以及在较小程度上的 IL-6 和 MCP-1 的表达也受到抑制。LPS 上调 Cyp27b1 基因表达，而 1.25（OH）2D3 部分逆转了这一表达。
这些数据表明维生素 D3（25（OH）D3 和 1.25（OH）2D3）对人巨噬细胞具有抗炎作用，并支持将高剂量维生素 D3 靶向免疫系统的方法可能对炎症条件具有有益的临床效果。
