中国兽医科学2024，Vol.54Issue(7)：974-981,8.DOI:10.16656/j.issn.1673-4696.2024.0118
siRNA干扰TLR4影响两种酵母多糖处理RAW264.7细胞NF-κB/MAPK信号通路的激活
Knockdown of TLR4 by siRNA affects the activation of NF-κB/MAPK signaling pathway in RAW264.7 cells treated with two kinds of zymosan
摘要
Abstract
In order to explore the role of Toll-like receptor 4(TLR4)in the activation of NF-κB/MAPK signaling pathway by two yeast polysaccharides.In this study,CCK8 method was used to detect the cytotoxicity of yeast β-glucan and mannan,and various methods were used to screen the best TLR4 siRNA.Then,ELISA and RT-qPCR were used to detect the secretion of related inflammatory factors and their mRNA expression.Finally,the phosphorylation level of key proteins in NF-κB/MAPK signaling pathway was detected by Western-blot.The results showed that 2.5-20 μg/mL β-glucan and 2.5-50 μg/mL mannan had no effect on the viability of RAW264.7 cells.TLR4 siRNA-2 can effectively knock down TLR4 mRNA and protein expression levels.The secretion of IL-6,IL-12,IL-1β,TNF-α,NO and the mRNA expression levels of IL-6,IL-12,IL-1β,TNF-α and iNOS were significantly decreased after TLR4 knockdown cells were treated with two yeast polysaccharides.The levels of key proteins p-P65,p-P38,p-ERK1/2,p-JNK and p-IκB in the NF-κB/MAPK signaling pathway were significantly decreased.The results showed that the two yeast polysaccharides could regulate the secretion of NF-κB/MAPK signaling pathway and related cytokines through TLR4,and finally determined that TLR4 was an important receptor for the immune regulation of the two yeast polysaccharides.关键词
酵母β-葡聚糖/酵母甘露聚糖/Toll样受体4/RAW264.7细胞/NF-κB/MAPK信号通路Key words
yeast β-glucan/yeast mannan/TLR4/RAW264.7 cells/NF-κB/MAPK signaling pathway分类
农业科技引用本文复制引用
杜蕊,陈灰,张宇飞,李松建,刘世雄,李薛强,徐子萱,薛晨,刘大程..siRNA干扰TLR4影响两种酵母多糖处理RAW264.7细胞NF-κB/MAPK信号通路的激活[J].中国兽医科学,2024,54(7):974-981,8.基金项目
国家自然科学基金项目(32060774) （32060774）
内蒙古自治区科技计划项目(2020GG0036 （）
2022YFDZ0051) （）
政府间国际科技创新合作项目(2023YFE0100400) （2023YFE0100400）
高等教育改革发展项目-研究生科研创新资助项目(S20231099Z) （S20231099Z）
