Department of Cell and Molecular Biology, School of Biology, College of Science, University of Tehran, P.O. Box: 141556455, Tehran, Iran.
Rheumatology Research Center, Shariati Hospital, Tehran University of Medical Sciences, Kargar Ave, P.O. Box: 1411713137, Tehran, Iran.
Sci Rep. 2021 Sep 8;11(1):17842. doi: 10.1038/s41598-021-96262-2.
Macrophages participate in the pathogenesis of ankylosing spondylitis (AS) by producing inflammatory cytokines. Extracellular adenosine triphosphate (eATP), released during cell stress, acts through purinergic receptors (P2XR and P2YR) and induces inflammatory responses. We investigated the effect of 2'(3')-O-(4-benzoyl benzoyl) ATP (BzATP) (a prototypic agonist of P2X7R) on the production of inflammatory cytokines in both monocyte-generated (M2-like) and M1 macrophages from patients and controls. Macrophages were differentiated from isolated periphery-monocytes (n = 14 in each group) by macrophage colony-stimulating factor (M-CSF). Using LPS and IFN-γ, macrophages were skewed toward M1 type and were treated with BzATP. Gene expression and protein release of IL-1β, IL-23, and TNF-α were evaluated by real-time PCR and ELISA methods respectively before and after treatment. BzATP significantly increased the protein release of TNF-α and the expression of TNFA and IL1B in monocyte-generated macrophages. Besides, BzATP treatment significantly upregulated IL1B expression, reduced TNFA and IL23A expression, and TNF-α release in M1 macrophages from both groups. Monocyte-generated and M1 macrophages from AS patients released higher TNF-α and expressed more IL1B in response to the same concentration of BzATP treatment respectively. Based on our results, AS macrophages were more sensitive to BzATP treatment and responded more intensively. Besides, the diverse effects of BzATP on monocyte-derived and M1 macrophages in our study may represent the differed inflammatory properties of these two groups of macrophages in response to eATP in the body.
巨噬细胞通过产生炎症细胞因子参与强直性脊柱炎 (AS) 的发病机制。细胞应激时释放的细胞外三磷酸腺苷 (eATP) 通过嘌呤能受体 (P2XR 和 P2YR) 发挥作用，诱导炎症反应。我们研究了 2'（3'）-O-（4-苯甲酰基苯甲酰基）三磷酸腺苷 (BzATP)（P2X7R 的典型激动剂）对患者和对照来源的单核细胞生成的 (M2 样) 和 M1 巨噬细胞中炎症细胞因子产生的影响。巨噬细胞通过巨噬细胞集落刺激因子 (M-CSF) 从分离的外周单核细胞中分化而来 (每组 n=14)。用 LPS 和 IFN-γ 将巨噬细胞向 M1 型倾斜，并给予 BzATP 处理。用实时 PCR 和 ELISA 方法分别在处理前后评估 IL-1β、IL-23 和 TNF-α 的基因表达和蛋白释放。BzATP 显著增加了单核细胞生成的巨噬细胞中 TNF-α 的蛋白释放和 TNFA 和 IL1B 的表达。此外，BzATP 处理还显著上调了两组 M1 巨噬细胞中 IL1B 的表达，降低了 TNFA 和 IL23A 的表达和 TNF-α 的释放。来自 AS 患者的单核细胞生成和 M1 巨噬细胞在对相同浓度的 BzATP 处理时分别释放更高水平的 TNF-α 和表达更多的 IL1B。根据我们的结果，AS 巨噬细胞对 BzATP 治疗更敏感，反应更强烈。此外，我们研究中 BzATP 对单核细胞来源的和 M1 巨噬细胞的不同作用可能代表了这两组巨噬细胞在体内对 eATP 反应的不同炎症特性。
