槲皮素对脂多糖刺激巨噬细胞炎症因子释放的影响

Effect of quercetin on LPS-stimulated macrophage inflammatory cytokine release

目的观察槲皮素对脂多糖（LPS）刺激巨噬细胞炎症因子释放的影响。方法采用四甲基偶氮唑盐微量酶反应比色（MTT）法测定不同浓度槲皮素对RAW264．7细胞活性的影响。将细胞分为五组：A组（对照组）、B组（LPS干预组）、C组（LPS＋槲皮素50μmol/L）、D组（LPS＋槲皮素150μmol／L）和E组（LPS＋槲皮素250μmol／L）。real—time测定各组细胞Toll样受体4（TLR4）、髓样分化因子88（Myd88）、NF—κB（p65）mRNA的表达；酶联免疫吸附测定（ELISA）测定各组细胞上清液白细胞介素6（IL-α）、肿瘤坏死因子-α（TNF—α）分泌水平。结果槲皮素在0-250μmol/L浓度范围内对RAW264-7细胞活性没有影响。同对照组相比，槲皮素可有效降低LPS引起的TLR4、MyD88、NF-κB（p65）的mRNA表达和细胞上清液中IL-α、TNF—α分泌水平，并呈剂量依赖性（P〈0．01）。结论槲皮素可能通过抑制TLR-Myd88-NF-κB传导通路的活化，减少巨噬细胞炎症因子的分泌。

Objective To study the impact of quercetin on LPS-stimulated macrophage inflammatory cytokine release. Methods Determined by MTT assay with different concentrations of quercetin on the activity of RAW 264.7 cells. Cells were divided into five groups： A （control group）, B group （LPS intervention group）, C （LPS＋quercetin 50 μmol/L）, D group （LPS＋quercetin 150μmol/L） and E group （LPS＋quercetin 250 μmol/L）. Cells expression of toll-like receptor 4 （TLR4） and myeloid differentiation factor 88 （Myd88） and NF-kappa B （p65） mRNA in each group was determined by real-time determination; Supernatant IL-α and TNF-α secretion level was tested by EL1SA determination. Results The quercetin at 0 to 250 μmol/L concentration range on the activity of RAW 264.7 cells did not significantly affect. Compared with the control group, quercetin can effectively reduce the level of LPS-induced TLR4 and of MyD88 and of NF-κB （p65） of mRNA expression and supematant IL-α and TNF-α secretion, and dose-dependent manner（P〈0.01）. Conclusion Quercetin may reduce macrophage inflammatory cytokine secretion by inhibiting the activation of TLR-Myd88-NF-κB pathway.