Cell Biology and Immunology Group, Wageningen University and Research Centre, Marijkeweg 40, P.O. Box 338, 6700 AH, Wageningen, The Netherlands.
Food Funct. 2010 Dec;1(3):254-61. doi: 10.1039/c0fo00113a. Epub 2010 Nov 1.
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-1β, IL-6, IL-8, IL-10 and TNF-α), inflammation-related enzymes (iNOS and COX-2), and transcription factors (NF-κB, 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-κB 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.
建立了一种分析方法，以研究食物化合物对 THP-1 细胞系来源的单核细胞和巨噬细胞中炎症相关免疫应答的影响。首先，我们集中研究了用脂多糖（LPS）或刀豆蛋白 A（ConA）刺激后的作用。采用 RT-PCR 分析了炎症相关细胞因子（IL-1β、IL-6、IL-8、IL-10 和 TNF-α）、炎症相关酶（iNOS 和 COX-2）和转录因子（NF-κB、AP-1 和 SP-1）的基因表达动力学。通过研究炎症相关细胞因子的蛋白水平分泌，来研究炎症相关反应。LPS 刺激诱导了 THP-1 单核细胞和 THP-1 巨噬细胞中炎症相关细胞因子、COX-2 和 NF-κB 基因的表达，最大上调分别发生在 3 小时和 6 小时。这些时间点随后被用于研究三种已知的免疫调节食物衍生化合物（槲皮素、柑橘果胶和大麦葡聚糖）的抗炎活性。LPS 与槲皮素、柑橘果胶或大麦葡聚糖共同刺激 THP-1 单核细胞和巨噬细胞显示出这些化合物的不同免疫调节活性。因此，我们提出，同时将 THP-1 细胞暴露于 LPS 和食物化合物中，并结合基因表达响应分析，是一种有前途的体外筛选工具，可以在有限的时间内选择具有抗炎作用的食物化合物。
