Transcription profiles of LPS-stimulated THP-1 monocytes and macrophages: a tool to study inflammation modulating effects of food-derived compounds
Transcription profiles of LPS-stimulated THP-1 monocytes and macrophages: a tool to study inflammation modulating effects of food-derived compounds
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DOI：
10.1039/c0fo00113a
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发表时间：
2010-12-01
期刊：
影响因子：
6.1
通讯作者：
Wichers, Harry J.
中科院分区：
文献类型：
--
作者：
Chanput, Wasaporn;Mes, Jurriaan;Wichers, Harry J.
建立了一种研究食物化合物对THP-1细胞来源的单核细胞和巨噬细胞的炎症相关免疫反应的方法。第一种策略侧重于内毒素(LPS)或刀豆蛋白A(ConA)刺激后的影响。RT-PCR分析炎症相关细胞因子(IL-1β、IL-6、IL-8、IL-10和TNF-α)、炎症相关酶(iNOS和COX-2)、转录因子(NF-kappa B、AP-1和SP-1)的基因表达动力学。通过时间依赖性细胞因子的分泌，从蛋白水平研究炎症反应。LPS刺激诱导THP-1单核细胞和THP-1巨噬细胞表达炎症相关细胞因子COX-2和NF-kappaB基因，分别在3h和6h表达上调。这些时间点随后被选择来研究三种众所周知的免疫调节食品衍生化合物的炎症调节活性：栎素、柑橘果胶和大麦葡聚糖。在THP-1单核细胞和巨噬细胞中，分别用槲皮素、柑橘果胶或大麦葡聚糖刺激脂多糖，显示出不同的免疫调节活性。因此，我们认为，同时将THP-1细胞暴露于脂多糖和食物化合物中，结合基因表达反应分析是一种有希望的体外筛选工具，可以在有限的时间框架内筛选出具有炎症调节作用的食物化合物。
An assay was developed to study inflammation-related immune responses of food compounds on monocytes and macrophages derived from THP-1 cell line. First strategy focused on the effects after stimulation with either lipopolysaccharide (LPS) or Concanavalin A (ConA). Gene expression kinetics of inflammation-related cytokines (IL-1 beta, IL-6, IL-8, IL-10 and TNF-alpha), inflammation-related enzymes (iNOS and COX-2), and transcription factors (NF-kappa B, AP-1 and SP-1) were analyzed using RT-PCR. Time dependent cytokine secretion was investigated to study the inflammation-related responses at protein level. LPS stimulation induced inflammation-related cytokine, COX-2 and NF-kappa B genes of THP-1 monocytes and THP-1 macrophages with the maximum up-regulation at 3 and 6 h, respectively. These time points, were subsequently selected to investigate inflammation modulating activity of three well known immuno-modulating food-derived compounds; quercetin, citrus pectin and barley glucan. Co-stimulation of LPS with either quercetin, citrus pectin, or barley glucan in THP-1 monocytes and macrophages showed different immuno-modulatory activity of these compounds. Therefore, we propose that simultaneously exposing THP-1 cells to LPS and food compounds, combined with gene expression response analysis are a promising in vitro screening tool to select, in a limited time frame, food compounds for inflammation modulating effects.