李恒,李锐,马景球.TLR4–MyD88信号转导途径介导仙人掌多糖免疫调节的机制研究[J].湖南农业大学学报:自然科学版,2021,47(4):.
TLR4–MyD88信号转导途径介导仙人掌多糖免疫调节的机制研究
  
DOI:
中文关键词:  仙人掌多糖  小鼠腹腔巨噬细胞  MyD88依赖性信号通路
英文关键词:Opuntia dillenii Haw. polysaccharides  mouse peritoneal macrophage  MyD88-dependent signaling pathway
基金项目:广东省自然科学基金项目(2016A030313669)
作者单位
李恒,李锐,马景球 岭南师范学院食品科学与工程学院广东 湛江 524048 
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中文摘要:
      为研究仙人掌多糖(ODPs)对Toll样受体4(TLR4)及其转导的MyD88依赖性信号途径中下游重要元件髓样分化因子88(MyD88)、肿瘤坏死因子受体相关因子6(TRAF6)、IκB激酶β(IKKβ) mRNA和蛋白表达的影响,体外培养小鼠腹腔单核巨噬细胞(RAW264.7),用ODPs干预24 h,收集细胞,分别采用RT–PCR法和蛋白质印迹Western blot法检测TLR4、MyD88、TRAF6、IKKβ mRNA及蛋白表达。结果显示:中、高剂量(100、200 μg/mL)ODPs显著增强TLR4、TRAF6及IKKβ的基因表达及TLR4、MyD88、TRAF6的蛋白表达(P<0.05),且具有良好的量效关系;ODPs也显著降低了MyD88的基因表达(P<0.05),对IKKβ的蛋白表达量增强效果不显著。说明非炎性条件下ODPs可促使TLR4高表达,同时激活TLR4胞内信号转导的MyD88依赖性途径。
英文摘要:
      To study the effect of different concentrations(50, 100, 200 μg/mL) of Opuntia dillenii Haw. polysaccharides (ODPs) on Toll-like receptor 4(TLR4) and MyD88-dependent signaling pathway, mouse macrophage cells(RAW264.7) were cultured in vitro and treated with ODPs for 24 hours first. Then, the cells were collected and real-time fluorescence quantitative RT-PCR was used to detect the gene expression of TLR4, MyD88, TRAF6 and IKKβ. Western blot was used to detect the expression of TLR4, MyD88, TRAF6 and IKKβ proteins. The results showed that the expression of TLR4, TRAF6 and IKKβ genes and the expression of TLR4, MyD88, and TRAF6 proteins were significantly increased by medium and high doses(100 and 200 μg/mL) of ODPs(P<0.05). They had a good dose-effect relationship. The expression of MyD88 gene was significantly reduced by different concentrations of ODPs(P<0.05). But the increase of IKKβ protein expression was not significant. In summary, ODPs can promote high expression of TLR4 gene and protein and activate the MyD88-dependent pathway of intracellular signal transduction of TLR4 under non-inflammatory conditions, which may be one of the mechanisms that ODPs exert their effects on enhancing immunity.
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