| 丛大鹏,郝杰杰,祝贺,等.一种 β-葡聚糖的结构表征及其体外免疫活性研究[J].中国海洋药物,2015,34(5):1-6. |
| 一种 β-葡聚糖的结构表征及其体外免疫活性研究 |
| Structural characterization and immune activity of a β-glucan from Durvillaea Antarctica |
| 投稿时间：2015-03-17 修订日期：2015-04-13 |
| DOI： |
| 中文关键词: 海茸 β-葡聚糖 结构表征 免疫活性 |
| English Keywords:Durvillaea Antarctica glucan structure characterization immune activity |
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| 中文摘要: |
| 目的 从海茸(Durvillaea antarctica)中提取、分离和纯化β-葡聚糖,并对其结构和体外免疫活性进行研究。方法 将干燥海茸粉碎,经水提取和Q-Sepharose Fast Flow离子交换层析分离得到β-葡聚糖(DAC)。采用硫酸-苯酚法、Folin-酚法、柱前衍生高效液相色谱法和多角度高效凝胶渗透色谱与多角度激光光散射器联用(HPGPC-MALLS)法对DAC的总糖含量、蛋白含量、单糖组成和分子量进行分析,并通过甲基化和气质联用(GC-MS)、傅里叶变换红外光谱(FTIR)和核磁共振波谱(NMR)对其结构进行表征。采用中性红法对DAC的体外免疫活性进行评价。结果 从海茸中分离纯化得到一种总糖含量达97.5%的β-葡聚糖,其蛋白含量和分子量分别为2.0 %和18.3 kDa。经甲基化和NMR分析结果表明,DAC是以→3)Glc(β1→为主链,并含有少量→6)Glc(α/β1→分支的葡聚糖。体外免疫活性结果显示,在浓度为50 μg.mL-1时, DAC对巨噬细胞的吞噬能力达到最大值。结论 海茸β-葡聚糖能显著增强巨噬细胞的吞噬能力,是一种潜在的免疫增强剂。 |
| English Summary: |
| Objective To extract and purify b-glucan from Durvillaea Antarctica, and investigate its structure and immune activity. Methods A water soluble glucan was obtained from Durvillaea Antarctica by water extraction at room temperature, and further purified by CaCl2 and strong anion exchange chromatography. The contents of total sugar and protein, the monosaccharide composition and the molecular mass of the glucan were analyzed by phenol-sulfuric method, folin-phenol method, precolumn derivatization reversed phase-high performance liquid chromatography (RP-HPLC) and high performance gel permeation chromatography – multi-angle laser light scattering (HPGPC-MALLS), respectively. The structure of the glucan was characterized based on fourier transform infra-red (FTIR) and nuclear magnetic resonance (NMR) spectroscopy, methylation analysis and gas chromatography-mass spectrometry (GC-MS) analysis. The immune activity was measured by neutral red method. Results The glucan from Durvillaea Antarctica (DAC) was composed of a backbone of→3)Glc(β1→ and a little branch of →6) Glc(α/β1→. The immune activity assay showed that 50 μg.mL-1 DAC could obviously increase the phagocytosis ability of mononuclear phagocytes in Raw 264.7 cell line. Conclusion The β-glucan from Durvillaea Antarctica had a significant immune activity in Raw 264.7 cell line, and could be used as a potential immunity intensifier. |
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