β-胡萝卜素的不同添加方式对脂多糖刺激的RAW264.7细胞炎症的影响

Effects of different β-carotene adding models on the inflammation of LPS-induced RAW264.7 cells

为探讨β-胡萝卜素不同添加方式(预防型和治疗型)对脂多糖(LPS)刺激的RAW264.7细胞的细胞活力、活性氧(ROS)、炎性因子以及NF-κB通路的影响,利用MTT法检测细胞活力确定β-胡萝卜素的最佳添加浓度和时间,流式细胞仪检测ROS的百分含量,荧光定量PCR检测炎性因子IL-1β、IL-6和TNF-αmRNA相对表达量;Western Blot测NF-κB p65蛋白的表达量分析β-胡萝卜素对NF-κB通路的影响。结果表明:对于LPS刺激的RAW264.7细胞,治疗型和预防型添加β-胡萝卜素均可以提高细胞活力,且在治疗型添加模型中,可极显著提高(P<0.01);治疗型和预防型添加β-胡萝卜素均可以降低ROS百分含量、炎性因子mRNA相对表达量和NF-κB p65蛋白表达量。综上所述,不同添加方式的β-胡萝卜素对LPS刺激的巨噬细胞的细胞活力均有提升作用,对ROS和炎性因子均有抑制作用,且这种抑制作用可能与NF-κB通路有关。

The aim of this study was to investigate the effects of β-carotene treatment model and prevention model on ceil viability,ROS,inflammatory factors and NF-β p65 protein in LPS-induced RAW264.7 cells. To determine the most suitable concentration and time of β-carotene,the cell viability in two models was measured by MTT. The percentage of ROS was detected by flow cytometry analysis, and the mRNA relative expressions of IL-1β, tL-6 and TNF-a were measured by fluorescence quantitative PCR. The protein relative expressions of NF-β p65 were measured by Western Blot to research the effect of β-carotene on NF-β signaling pathway. The results showed that.ln LPS-induced RAW264.7 cells, the cell viabilities of treatment model and prevention model were higher and the difference was very significant in treatment model （P〈0.01）;The percentage of ROS, expression of inflammatory factors and protein NF-B p65 were lower than LPS-induced group. In summary,β-carotene promoted the cell viability of LPS-induced RAW264.7 cells and inhibited the percentage of ROS and expression of inflammatory factors,indicating the correlation of the inhibition effect with NF-B pathway.