[关键词]
[摘要]
目的 从何首乌Polygonum multiflorum中提取分离多糖,对其结构进行表征,并评价其免疫调节活性。方法 采用水提、醇沉法从何首乌中提取粗多糖,经Q-Sepharose Fast Flow强阴离子交换色谱柱分离纯化得到何首乌多糖(PMT)。利用高效凝胶渗透色谱-十八角度激光光散射联用法(HPGPC-MALLS)测定PMT的绝对分子质量,采用PMP柱前衍生-高效液相色谱法(HPLC)测定单糖组成。采用二维核磁共振波谱(2D-NMR)对PMT的结构进行表征。采用四甲偶氮唑盐(MTT)法、中性红比色法和Griess法分别检测PMT对小鼠单核巨噬细胞RAW264.7的生长、吞噬活性和释放一氧化氮(NO)能力的影响。结果 PMT是一种α-1,4-葡聚糖,其绝对分子质量为3.96×105。PMT可显著促进RAW264.7细胞的增殖,促进细胞的吞噬能力,增加NO的释放量。结论 何首乌来源的α-1,4-葡聚糖具有显著的免疫调节活性,具有药物开发潜力。
[Key word]
[Abstract]
Objective To extract and separate a polysaccharide from Polygonum multiflorum, characterize its structural features and study its immunomodulatory activity. Methods The polysaccharide from P. multiflorum (PMT) was isolated and purified by water extraction and ethanol precipitation following Q-Sepharose Fast Flow ion exchange chromatography column. Molecular weight of PMT was determined by high performance gel permeation chromatography-multiple angle laser light scattering (HPGPC-MALLS), and monosaccharide composition was analyzed by HPLC with PMP (1-phenyl-3-methyl-5-pyrazolone) pre-column derivatization, respectively. The structure of PMT was characterized by proton nuclear magnetic resonance spectrum (2D-NMR). The immunomodulatory activities were tested by MTT, neutral red colorimetric assay and Griess method. Results PMT was a kind of α-1,4-glucan, and its molecular mass was 3.96×105. PMT promoted the proliferation and phagocytosis of RAW 264.7 cells, and significantly induced the increase of NO production in a dose-dependent manner. Conclusion The polysaccharide from P. multiflorum is a linear α-1,4-glucan with potent immunomodulatory activity, which would be potentially developed as an effective drug.
[中图分类号]
R284.1
[基金项目]