摘要: |
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-α mRNA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。 |
关键词: β-胡萝卜素 RAW264.7细胞 细胞活力 活性氧 炎性因子 |
DOI:10.11841/j.issn.1007-4333.2017.11.13 |
投稿时间:2016-11-25 |
基金项目:国家自然科学基金面上项目(31672511)。 |
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Effects of different β-carotene adding models on the inflammation of LPS-induced RAW264.7 cells |
ZHANG Xiaoyin, WU Min, ZHANG Shanshan, JI Yubin, HUANG Yusheng, ZHENG Xin
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(College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China) |
Abstract: |
The aim of this study was to investigate the effects of β-carotene treatment model and prevention model on cell viability,ROS,inflammatory factors and NF-κ B 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-1 β, IL-6 and TNF-α were measured by fluorescence quantitative PCR.The protein relative expressions of NF-κ B p65 were measured by Western Blot to research the effect of β-carotene on NF-κ B signaling pathway.The results showed that: In 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-κ 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-κ B pathway. |
Key words: β-carotene RAW264.7 cell cell viability ROS inflammatory factor |