School of Biotechnology and Food Engineering, Hefei University of Technology, PR China.
School of Biotechnology and Food Engineering, Hefei University of Technology, PR China.
Carbohydr Polym. 2015 Dec 10;134:66-73. doi: 10.1016/j.carbpol.2015.07.070. Epub 2015 Jul 26.
In this work, a novel water-soluble homogeneous polysaccharide (LJP-31) with a molecular mass of 2.24 × 10(6) Da was isolated and purified from Laminaria japonica using DEAE-cellulose and Sephacryl S500 chromatography. Results showed that LJP-31 mainly consists of arabinose, mannose, glucose and galactose in a molar ratio of 1.0:7.8:6.6:0.8. LJP-31 exhibited significant stimulation on macrophages and enhanced the production of NO, TNF-α, IL-1β, IL-6 and IL-10 as well as the up-regulation of their gene expressions. Western blot analysis suggested that LJP-31 has the positive effects on the translocation of NF-κB p65 from cytoplasm to nucleus and the phosphorylation of IκBα, ERK1/2, JNK1/2 and P38 in macrophages. Flow cytometric and confocal laser-scanning microscopy analysis indicated that toll-like receptor 4 (TLR4) was at least one of the recognition receptors of LJP-31 on the plasma membrane of macrophages. Taken together, LJP-31 may exert its immunostimulating potency via TLR4 activation of MAPK and NF-κB signaling pathways.
在这项工作中,从海带中分离纯化出一种新型水溶性均一多糖(LJP-31),其分子量为 2.24×10(6)Da。采用 DEAE-纤维素和 Sephacryl S500 层析法对其进行分离纯化。结果表明,LJP-31 主要由阿拉伯糖、甘露糖、葡萄糖和半乳糖组成,摩尔比为 1.0:7.8:6.6:0.8。LJP-31 对巨噬细胞有明显的刺激作用,能增强 NO、TNF-α、IL-1β、IL-6 和 IL-10 的产生,并上调其基因表达。Western blot 分析表明,LJP-31 能促进 NF-κB p65 从胞浆向核内转移,以及 IκBα、ERK1/2、JNK1/2 和 P38 的磷酸化。流式细胞术和共聚焦激光扫描显微镜分析表明,Toll 样受体 4(TLR4)是 LJP-31 识别巨噬细胞膜上的受体之一。综上所述,LJP-31 可能通过 TLR4 激活 MAPK 和 NF-κB 信号通路发挥其免疫刺激作用。