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24h,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、IKKβmRNA及蛋白表达。结果显示:中、高剂量(100、200μg/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。
To study the effect of different concentrations(50,100,200μg/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 IKKβproteins.The results showed that the expression of TLR4,TRAF6 and IKKβgenes and the expression of TLR4,MyD88,and TRAF6 proteins were significantly increased by medium and high doses(100 and 200μg/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 IKKβprotein 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.
作者
李恒
李锐
马景球
LI Heng;LI Rui;MA Jingqiu(College of Food Science and Engineering,Lingnan Normal University,Zhanjiang,Guangdong 524048,China)
出处
《湖南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2021年第4期419-423,共5页
Journal of Hunan Agricultural University(Natural Sciences)
基金
广东省自然科学基金项目(2016A030313669)。
关键词
仙人掌多糖
小鼠腹腔巨噬细胞
MyD88依赖性信号通路
Opuntia dillenii Haw.polysaccharides
mouse peritoneal macrophage
MyD88-dependent signaling pathway
