槲皮素调控锌稳态抑制脂多糖诱导的小鼠巨噬细胞炎症反应

Quercetin inhibiting lipopolysaccharide-induced inflammation in mouse macrophages by modulating zinc homeostasis

目的:探讨槲皮素(QUE)对脂多糖(LPS)诱导的小鼠巨噬细胞炎症反应的作用及其机制。方法:将小鼠RAW264.7巨噬细胞分为正常对照组、LPS组和QUE+LPS组。采用酶联免疫吸附试验(ELISA)法检测细胞培养上清液中白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α和IL-6水平,流式细胞仪检测FluoZin-3标记细胞内游离锌含量,实时荧光定量PCR(RT-qPCR)检测各组细胞中ZIP8 mRNA表达,western blotting法检测ZIP8蛋白表达。结果:与LPS组(1μg/mL)相比,QUE+LPS组上清液中IL-1β、TNF-α和IL-6含量降低(P<0.01);与LPS组相比,QUE+LPS组巨噬细胞内游离锌含量降低(P<0.05),并可抑制LPS诱导的ZIP8表达(P<0.05)。结论:QUE可抑制LPS诱导的巨噬细胞炎症反应,其作用可能与下调ZIP8表达,抑制锌内流,减少巨噬细胞内游离锌含量有关。

Objective:To investigate the effect and mechanism of quercetin(QUE)on lipopolysaccharide(LPS)-induced inflammatory response in macrophages of mice.Methods:Mouse RAW264.7 macrophages were divid-ed into normal control group,LPS group,and QUE+LPS group.The levels of interleukin(IL)-1β,tumor necrosis factor(TNF)-αand IL-6 in cell culture supernatants were detected by enzyme-linked immunosorbent assay(ELI-SA).The free zinc content in FluoZin-3 labeled cells was detected by flow cytometry,the levels of ZIP8 mRNA expression in each group of cells were detected by real-time fluorescence quantitative PCR(RT-qPCR),and the expression of ZIP8 protein was detected by western blotting.Results:Compared with the LPS group(1μg/mL),the contents of IL-1β,TNF-αand IL-6 in the macrophage supernatants of the QUE+LPS group were decreased(P<0.01).Compared with the LPS group,the content of free zinc in macrophages of the QUE+LPS group were decreased(P<0.05)and the LPS-induced ZIP8 expression were inhibited(P<0.05).Conclusion:QUE can in-hibit LPS-induced macrophage inflammatory response,and its effect may be related to down-regulating ZIP8 ex-pression,inhibiting zinc influx,and reducing free zinc content in macrophages.