摘要
探究肉苁蓉低分子糖对RAW264.7小鼠巨噬细胞的激活作用及潜在机制。该实验将RAW264.7细胞分为正常对照组、细菌脂多糖(LPS)阳性对照组、肉苁蓉低分子糖处理组,其中肉苁蓉低分子糖的给药浓度为3.91~62.5 g·L-1。采用中性红法检测巨噬细胞吞噬活性,一氧化氮(NO)试剂盒检测NO的释放量,Western blot法检测巨噬细胞激活相关蛋白(TNF-α,IL-6,IKKβ,p-IKKβ,IκBα,p-IκBα,NF-κB,p-NF-κB)的表达。结果显示,肉苁蓉低分子糖对巨噬细胞具有激活作用,具体表现在:其可显著提高RAW264.7细胞NO的释放量,并促进细胞因子TNF-α和IL-6的表达。同时,其还可显著提高NF-κB的磷酸化及其上游关键信号蛋白IKKβ和IκBα的磷酸化水平。另外,低分子糖中的甘露醇可以使巨噬细胞的吞噬活性显著提升。以上结果表明,肉苁蓉低分子糖对巨噬细胞具有激活作用,其潜在机制是通过NF-κB信号通路来实现的,甘露醇可能是其中的一种关键活性单体成分。
To investigate the immune activation effect and mechanism of low molecular weight saccharides from Cistanche deserticola （LMSC） on mouse peritoneal macrophages, RAW264. 7 cells. The RAW264. 7 cells were divided into the normal control group, LPS positive control group, and LMSC treatment groups. The RAW264. 7 cells were treated with various concentrations of LMSC from 3.91 to 62. 5 g L-1. The neutral red assay was employed to detect the phagocytic activity of macrophages. NO release was detected by u- sing NO kit, and macrophage activation associated protein expression levels （TNF-α, IL-6, IKK/3, p-IKK/3, IkBα, p-IkBα, NF-KB, and p-NF-kB） were detected by Western blot. Results showed that LMSC had an activation effect on macrophages; it can significantly increase the release of NO in RAW264. 7 ceils and promote the expression of cytokines TNF-α and IL-6. Moreover, LMSC significantly increased the phosphorylation of IKK/3, IkBα, and NF-kB p65. Furthermore, mannitol＇s one of the main constituents in LMSC signifi- cantly enhanced the phagocytic activity of macrophages. These results showed that LMSC could activate macrophages by up-regulating the NF-t^B signaling pathway, and mannitol may be one of the main active components in LMSC.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2017年第21期4207-4210,共4页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81303253)
关键词
肉苁蓉低分子糖
免疫激活
巨噬细胞
炎症因子
NF-ΚB
Low molecular weight saccharides
Cistanche deserticola
immune activation
macrophage
inflammatory, factor
NF-kB
