Vitamin D3 Metabolites Enhance the NLRP3-Dependent Secretion of IL-1β From Human THP-1 Monocytic Cells
Vitamin D3 Metabolites Enhance the NLRP3-Dependent Secretion of IL-1β From Human THP-1 Monocytic Cells
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DOI：
10.1002/jcb.24985
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发表时间：
2015-05-01
影响因子：
4
通讯作者：
MacDonald, Justin A.
中科院分区：
文献类型：
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作者：
Tulk, Sarah E.;Liao, Kuo-Chieh;MacDonald, Justin A.
维生素 D-3 已成为免疫系统的重要调节剂。由于目前在多种免疫细胞中发现了维生素 D-3 活化代谢酶和维生素 D 受体 (VDR)，活性维生素 D-3 代谢物 1,25(OH)(2)D-3 被认为具有免疫调节特性。我们检查了 1,25(OH)(2)D-3 是否也可能增强巨噬细胞依赖 NLRP3 释放成熟 IL-1 β。用维生素 D-3 代谢物刺激 PMA 分化的 THP-1 细胞，并通过蛋白质印迹和实时 qPCR 评估 CYP27、CYP24、NLRP3、ASC、procaspase-1 表达，并通过 ELISA 测量促炎细胞因子 IL-1b 释放来评估炎症小体激活。暴露于1,25(OH)(2)D-3对VDR的基础表达水平没有影响；然而，CYP27A1 转录本受到抑制，而 CYP24A1 转录本则大幅升高。 1,25(OH)(2)D-3- 和 25(OH) D-3 均诱导 THP-1 细胞释放 IL-1b，并且使用 caspase-1 抑制剂 YVAD 和 NLRP3 抑制剂格列本脲和 Bay 11-7082 可以阻断这些作用。有趣的是，1,25(OH)(2)D-3 暴露降低了 NLRP3 蛋白表达，但对 ASC 或 pro-caspase-1 蛋白水平没有影响。与 ATP 或艰难梭菌毒素相比，1,25(OH)(2)D-3 引起的成熟 IL-1b 的增加是适度的。然而，与单独使用 ATP 或 1,25(OH)(2)D-3 相比，用 ATP 和 1,25(OH)(2)D-3 共同处理 THP-1 细胞会导致更多的 IL-1b 分泌。 J.细胞。生物化学。 116：711-720，2015。(C)2015 Wiley 期刊公司。
Vitamin D-3 has emerged as an important regulator of the immune system. With metabolic enzymes for vitamin D-3 activation and vitamin D receptors (VDR) now identified in a variety of immune cells, the active vitamin D-3 metabolite 1,25(OH)(2)D-3, is thought to possess immunomodulatory properties. We examined whether 1,25(OH)(2)D-3 might also enhance the NLRP3-dependent release of mature IL-1 beta from macrophages. PMA-differentiated THP-1 cells were stimulated with vitamin D-3 metabolites and assessed for CYP27, CYP24, NLRP3, ASC, procaspase-1 expression by western blot and real-time qPCR as well as inflammasome activation with pro-inflammatory cytokine IL-1b release measured by ELISA. Exposure to 1,25(OH)(2)D-3 had no effect on the basal expression levels of VDR; however, CYP27A1 transcript was suppressed and CYP24A1 transcript was substantively elevated. Both 1,25(OH)(2)D-3- and 25(OH) D-3 induced IL-1b release from THP-1 cells, and these effects were blocked with application of the caspase-1 inhibitor YVAD and the NLRP3 inhibitors glyburide and Bay 11-7082. Interestingly, 1,25(OH)(2)D-3 exposure reduced NLRP3 protein expression but had no effect on ASC or pro-caspase-1 protein levels. The increase in mature IL-1b elicited by 1,25(OH)(2)D-3 was modest compared to that found for ATP or C. difficile toxins. However, co-treatment of THP-1 cells with ATP and 1,25(OH)(2)D-3 resulted in more IL-1b secretion than ATP or 1,25(OH)(2)D-3 alone. J. Cell. Biochem. 116: 711-720, 2015. (C) 2015 Wiley Periodicals, Inc.