Wang Yang, Shen Xiaoyan, Yin Kaiyue, Miao Changqing, Sun Yanlong, Mao Shumei, Liu Dongmei, Sheng Jiwen
Department of Pharmacy, Weifang Medical University, Weifang 261053, China.
Department of Pharmacy, Weifang Medical University, Weifang 261053, China.
Int J Biol Macromol. 2022 Apr 30;205:76-89. doi: 10.1016/j.ijbiomac.2022.02.037. Epub 2022 Feb 15.
Polysaccharides coded as CP were extracted from Athyrium Multidentatum (Doll.) Ching and then fractionated into five fractions (FP-1, FP-2, FP-3, FP-4 and FP-5). A purified polysaccharide designated as FP-3-4 was prepared from FP-3 by Sephadex G-100 column chromatography. Chemical analysis disclosed that CP and these fractions were heteropolysaccharides and mainly composed of glucose, galactose, arabinose, mannose, rhamnose, xylose, fucose, ribose and uronic acid with different molar ratios. They presented different images of SEM. FP-3-4 was highly branched polymers with sixteen types of linkages. The in vitro immunomodulatory results stated that CP and these fractions could promote macrophage proliferation, enhance macrophage phagocytosis and increase the production of NO, TNF-α, IFN-γ, IL-1β, IL-6, IL-10 and IL-2, indicating remarkable immune enhancement activities. RNA sequencing analysis revealed that CP and FP-3 induced macrophage activation mainly through MAPK and alternative NF-κΒ signaling pathways via CD14/TLR4 and Dectin-2 receptors, which were verified by RT-qPCR and western blot.
从多齿蹄盖蕨（Athyrium Multidentatum (Doll.) Ching）中提取编码为CP的多糖，然后将其分离为五个级分（FP-1、FP-2、FP-3、FP-4和FP-5）。通过葡聚糖G-100柱色谱法从FP-3制备了一种命名为FP-3-4的纯化多糖。化学分析表明，CP和这些级分是杂多糖，主要由葡萄糖、半乳糖、阿拉伯糖、甘露糖、鼠李糖、木糖、岩藻糖、核糖和糖醛酸组成，其摩尔比不同。它们呈现出不同的扫描电子显微镜图像。FP-3-4是具有十六种连接类型的高度分支聚合物。体外免疫调节结果表明，CP和这些级分可促进巨噬细胞增殖，增强巨噬细胞吞噬作用，并增加NO、TNF-α、IFN-γ、IL-1β、IL-6、IL-10和IL-2的产生，表明具有显著的免疫增强活性。RNA测序分析显示，CP和FP-3主要通过CD14/TLR4和Dectin-2受体经由MAPK和替代性NF-κB信号通路诱导巨噬细胞活化，这通过RT-qPCR和蛋白质印迹得到验证。
