阿里红多糖对RAW264.7巨噬细胞中NF-κB信号途径及肿瘤坏死因子-α表达的影响
目的 研究阿里红粗多糖(Fomes Officinals polysaccharide, FOPS)及阿里红多糖组分(FOPS-a)对巨噬细胞RAW264.7的Toll样受体4(toll-like receptor 4, TLR4)、核转录因子-κb p65(nuclear factor-κB p65, NF-κB p65)表达以及肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)、白介素-1β(interleukin-1β, IL-1β)、白介素-6(interleukin-6, IL-6)释放的影响, 初步探讨阿里红多糖组分的免疫调节机制。方法 采用RT-PCR法检测不同浓度(50、100、200 μg/mL)的FOPS及FOPS-a对IL-1β、TNF-α、p65、TLR4 mRNA表达量的影响; 用Western Blot法检测FOPS及FOPS-a对磷酸化核因子-κB(p-NF-κBp65)和磷酸化核因子抑制蛋白(p-Iκbα)表达水平的影响; 用NF-κB抑制剂BAY11-7082预处理细胞后, ELISA法检测上清液中TNF-α、IL-1β、IL-6分泌量的影响。结果 与空白对照组相比, 不同浓度的阿里红多糖可以显著提高TLR4、p65、TNF-α、IL-1β mRNA表达量和p-NF-κBp65和p-Iκbα的磷酸化水平(P<0.05); 通过阻断NF-κB途径可以显著抑制FOPS及FOPS-a对TNF-α、IL-1β、IL-6促进分泌的作用(P<0.05)。结论 FOPS及FOPS-a发挥免疫调节作用的机制与其激活NF-κB信号途径进而调节TNF-α等因子的基因表达和控制细胞因子释放有关。
Objective To investigate the effect of crude Fomes Officinals polysaccharide (FOPS) and its components on expressions of toll-like receptor 4 (TLR4) and nuclear factor-κB p65 (NF-κB p65), and release of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6)of RAW264.7 macrophages, and to preliminarily explore the immunomodulatory mechanism of Fomes Officinals polysaccharide components. Methods RT-PCR was used to detect mRNA expression levels of IL-1β, TNF-α, p65 and TLR4 in different concentrations (50, 100, 200 μg/mL) of FOPS and FOPS-a. Western Blot was used to detect the expression levels of phosphorylated nuclear factor-κB (p-NF-κBp65) and phosphorylated nuclear factor inhibitory protein (p-Iκbα) by FOPS and FOPS-a. After pretreatment of cells with NF-κB inhibitor BAY11-7082, the effects of TNF-α, IL-1β and IL-6 secretion in supernatant were detected by ELISA. Results Compared with the blank control group, Fomes Officinals at different concentrations could significantly increase the mRNA expression levels of TLR4, p65, TNF-α, and IL-1β and the phosphorylation levels of p-NF-κBp65 and p-Iκbα (P<0.05). Blocking NF-κB pathway could significantly inhibit the secretion of FOPS and FOPS-a on TNF-α, IL-1β and IL-6 (P<0.05). Conclusion The mechanism of immunomodulatory effect of FOPS and FOPS-a was related to their activation of NF-κB signaling pathway, thus regulating the gene expression of TNF-α and controlling the release of cytokines.
标题：阿里红多糖对RAW264.7巨噬细胞中NF-κB信号途径及肿瘤坏死因子-α表达的影响
英文标题：Effects of Fomes Officinals polysaccharide on NF-κB signaling pathway and expression of tumor necrosis factor-α in RAW264.7 cell
作者：
木尼萨.迪力夏提 新疆医科大学药学院
米仁沙.牙库甫 新疆医科大学药学院
伊明.尕哈甫 新疆医科大学药学院
丛媛媛 新疆医科大学药学院
帕丽达.阿不力孜 新疆医科大学药学院
中文关键词:阿里红多糖,阿里红多糖组分-a,巨噬细胞RAW264.7,核转录因子-κB,肿瘤坏死因子-α,
英文关键词:Fomes Officinals polysaccharide,Fomes officinalis polysaccharides-a,RAW264.7 cell,nuclear factor-κB,tumor nerosis factor-α,
发表日期：2018-10-30
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