High Uric Acid Activates the ROS-AMPK Pathway, Impairs CD68 Expression and Inhibits OxLDL-Induced Foam-Cell Formation in a Human Monocytic Cell Line, THP-1
High Uric Acid Activates the ROS-AMPK Pathway, Impairs CD68 Expression and Inhibits OxLDL-Induced Foam-Cell Formation in a Human Monocytic Cell Line, THP-1
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高尿酸激活 ROS-AMPK 通路，损害 CD68 表达并抑制人单核细胞系 THP-1 中 OxLDL 诱导的泡沫细胞形成。
DOI：
10.1159/000452567
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发表时间：
2016-01-01
影响因子：
--
通讯作者：
Cheng, Jidong
中科院分区：
文献类型：
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作者：
Luo, Chaohuan;Lian, Xueke;Cheng, Jidong
背景/目标：高尿酸血症是腹部肥胖、糖耐量异常、胰岛素抵抗、血脂异常和高血压等代谢综合征群的一部分。单核细胞/巨噬细胞在代谢综合征（包括痛风、肥胖和动脉粥样硬化）的发展中至关重要。然而，高尿酸（HUA）暴露如何影响单核细胞/巨噬细胞功能仍不清楚。在这项研究中，我们研究了单核细胞/巨噬细胞中HUA暴露的分子机制及其对氧化低密度脂蛋白（oxLDL）诱导的人单核细胞系THP-1泡沫细胞形成的影响。研究方法：我们用佛波醇-12-肉豆蔻酸酯-13-乙酸酯（PMA）诱导THP-1细胞分化，然后将细胞暴露于HUA，检测活性氧（ROS）的产生并分析磷酸化AMPK α的水平。在PMA之前，将THP-1细胞与化合物C（AMPK抑制剂）或N-乙酰基-L-半胱氨酸（NAC）（ROS清除剂）或HUA预孵育，以评估CD 68表达和磷酸-AMPKa水平。在化合物C和HUA处理之前，用oxLDL预处理PMA致敏的THP-1细胞。Western blot检测AMPKa磷酸化、CD 68、ABCG 1、ABCA 1、环氧化酶2（考克斯-2）和NF-κ B（p65）的表达。流式细胞术用于评估活细胞中的ROS产生和CD 68表达。油红0染色观察细胞摄取oxLDL的情况。结果：HUA处理增加PMA致敏的THP-1细胞中ROS的产生; NAC阻断HUA诱导的氧化应激。HUA处理时间依赖性地增加PMA引发的THP-1细胞中磷酸化AMPK α水平。HUA诱导的氧化应激增加磷酸化AMPK α水平，这被NAC阻断。HUA处理通过激活AMPK途径损害细胞分化期间的CD 68表达，这被化合物C处理逆转。最后，HUA处理抑制了THP-1细胞中泡沫细胞形成中的oxLDL摄取，这被化合物C处理阻断。HUA处理显著增加ABCG 1的表达并逆转oxLDL降低的ABCG 1表达，但不影响ABCA 1、NE-κ B（p65）或考克斯-2的表达。结论：HUA暴露激活ROS-AMPK通路，损害CD 68表达，并抑制oxLDL诱导的人单核细胞系THP-1泡沫细胞形成。(C)2016年，由S。Karger AG，巴塞尔。
Background/Aims: Hyperuricemia is part of the metabolic-syndrome cluster of abdominal obesity, impaired glucose tolerance, insulin resistance, dyslipidemia, and hypertension. Monocytes/macrophages are critical in the development of metabolic syndrome, including gout, obesity and atherosclerosis. However, how high uric acid (HUA) exposure affects monocyte/macrophage function remains unclear. In this study, we investigated the molecular mechanism of HUA exposure in monocytes/macrophages and its impact on oxidized low density lipoprotein (oxLDL)-induced foam-cell formation in a human monocytic cell line, THP-1. Methods: We primed THP-1 cells with phorbol-12-myristate-13-acetate (PMA) for differentiation, then exposed cells to HUA and detected the production of reactive oxygen species (ROS) and analyzed the level of phospho-AMPK alpha. THP-1 cells were pre-incubated with Compound C, an AM PK inhibitor, or N-acetyl-L-cysteine (NAC), a ROS scavenger, or HUA before PMA, to assess CD68 expression and phospho-AMPKa level. PMA-primed THP-1 cells were pre-treated with oxLDL before Compound C and HUA treatment. Western blot analysis was used to examine the levels of phospho-AMPKa, CD68, ABCG1, ABCA1, cyclooxygenase-2 (COX-2) and NF-KB (p65). Flow cytometry was used to assess ROS production and CD68 expression in live cells. Oil-red 0 staining was used to observe oxLDL uptake in cells. Results: HUA treatment increased ROS production in PMA-primed THP-1 cells; NAC blocked HUA-induced oxidative stress. HUA treatment time-dependently increased phospho-AMPK alpha level in PMA-primed THP-1 cells. The HUA-induced oxidative stress increased phospho-AMPK alpha levels, which was blocked by NAC. HUA treatment impaired CD68 expression during cell differentiation by activating the AMPK pathway, which was reversed by Compound C treatment. Finally, HUA treatment inhibited oxLDL uptake in the formation of foam cells in THP-1 cells, which was blocked by Compound C treatment. HUA treatment significantly increased the expression of ABCG1 and reversed the oxLDL-reduced ABCG1 expression but did not affect the expression of ABCA1, NE-kappa B (p65) or COX-2. Conclusions: HUA exposure activated the ROS-AMPK pathway, impaired CD68 expression, and inhibited oxLDL-induced foam-cell formation in a human monocytic cell line, THP-1. (C) 2016 The Authorts) Published by S. Karger AG, Basel.