Abstract：
Hyperuricemia has been shown to be an inducer of pro-inflammatory mediators by human primary monocytes. To study the deleterious effects of hyperuricemia, a reliable and stable in vitro model using soluble urate is needed. One recent report showed different urate-dissolving methods resulted in either pro-inflammatory or anti-inflammatory properties. The aim of this study was to compare the effect of two methods of dissolving urate on both primary human peripheral blood mononuclear cells (PBMCs) and THP-1 cells. The two methods tested were \'pre-warming\' and \'dissolving with NaOH\'.
Primary human PBMCs and THP-1 cells were exposed to urate solutions, prepared using the two methodologies: pre-warming and dissolving with NaOH. Afterwards, cells were stimulated with various stimuli, followed by the measurement of the inflammatory mediators IL-1β, IL-6, IL-1Ra, TNF, IL-8, and MCP-1.
In PBMCs, we observed an overall pro-inflammatory effect of urate, both in the pre-warming and the NaOH dissolving method. A similar pro-inflammatory effect was seen in THP-1 cells for both dissolving methods after restimulation. However, THP-1 cells exhibited pro-inflammatory profile with exposure to urate alone without restimulation. We did not find MSU crystals in our cellular assays.
Overall, the urate dissolving methods do not have critical impact on its inflammatory properties. Soluble urate prepared using either of the two methods showed mostly pro-inflammatory effects on human primary PBMCs and monocytic cell line THP-1. However, human primary PBMCs and the THP-1 differ in their response to soluble urate without restimulation.
Primary human PBMCs and THP-1 cells were exposed to urate solutions, prepared using the two methodologies: pre-warming and dissolving with NaOH. Afterwards, cells were stimulated with various stimuli, followed by the measurement of the inflammatory mediators IL-1β, IL-6, IL-1Ra, TNF, IL-8, and MCP-1.
In PBMCs, we observed an overall pro-inflammatory effect of urate, both in the pre-warming and the NaOH dissolving method. A similar pro-inflammatory effect was seen in THP-1 cells for both dissolving methods after restimulation. However, THP-1 cells exhibited pro-inflammatory profile with exposure to urate alone without restimulation. We did not find MSU crystals in our cellular assays.
Overall, the urate dissolving methods do not have critical impact on its inflammatory properties. Soluble urate prepared using either of the two methods showed mostly pro-inflammatory effects on human primary PBMCs and monocytic cell line THP-1. However, human primary PBMCs and the THP-1 differ in their response to soluble urate without restimulation.
摘要：
背景:已经证明高尿酸血症是人原代单核细胞的促炎介质的诱导物。为了研究高尿酸血症的有害影响，需要使用可溶性尿酸盐的可靠和稳定的体外模型。最近的一份报告显示不同的尿酸盐溶解方法导致促炎或抗炎特性。这项研究的目的是比较两种溶解尿酸盐的方法对原代人外周血单核细胞（PBMC）和THP-1细胞的影响。测试的两种方法是“预热”和“用NaOH溶解”。
方法：将原代人PBMC和THP-1细胞暴露于尿酸盐溶液，使用两种方法制备:预热和用NaOH溶解。之后,细胞被各种刺激刺激，然后测量炎症介质IL-1β，IL-6,IL-1Ra,TNF,IL-8和MCP-1。
结果：在PBMC中，我们观察到尿酸盐的整体促炎作用，在预热和NaOH溶解方法中。对于再刺激后的两种溶解方法,在THP-1细胞中观察到相似的促炎作用。然而,THP-1细胞在暴露于单独的尿酸盐而没有再刺激的情况下表现出促炎谱。在我们的细胞测定中我们没有发现MSU晶体。
结论：总体而言，尿酸盐溶解方法对其炎症特性没有关键影响。使用两种方法中的任一种制备的可溶性尿酸盐显示出对人类原代PBMC和单核细胞系THP-1的主要促炎作用。然而,人原发性PBMC和THP-1对可溶性尿酸盐的反应不同，而没有再刺激。
方法：将原代人PBMC和THP-1细胞暴露于尿酸盐溶液，使用两种方法制备:预热和用NaOH溶解。之后,细胞被各种刺激刺激，然后测量炎症介质IL-1β，IL-6,IL-1Ra,TNF,IL-8和MCP-1。
结果：在PBMC中，我们观察到尿酸盐的整体促炎作用，在预热和NaOH溶解方法中。对于再刺激后的两种溶解方法,在THP-1细胞中观察到相似的促炎作用。然而,THP-1细胞在暴露于单独的尿酸盐而没有再刺激的情况下表现出促炎谱。在我们的细胞测定中我们没有发现MSU晶体。
结论：总体而言，尿酸盐溶解方法对其炎症特性没有关键影响。使用两种方法中的任一种制备的可溶性尿酸盐显示出对人类原代PBMC和单核细胞系THP-1的主要促炎作用。然而,人原发性PBMC和THP-1对可溶性尿酸盐的反应不同，而没有再刺激。
