华东师范大学学报(自然科学版) ›› 2010, Vol. 2010 ›› Issue (4): 92-102.

• 生命科学 • 上一篇    下一篇

灰树花多糖硫酸酯化衍生物的制备与抑制HMEC迁移的作用

毛逸嵘， 张 易， 张红霞， 翁 亮， 张红锋   

  1. 华东师范大学 生命科学学院，上海 200062
  • 收稿日期:2009-04-01 修回日期:2009-08-01 出版日期:2010-07-25 发布日期:2010-07-25
  • 通讯作者: 张红锋

Preparation of Grifola frondosa polysaccharide sulfatedderivatives and their inhibitory effects on HMEC migration

MAO Yi-rong, ZHANG Yi, ZHANG Hong-xia, WENG Liang, ZHANG Hong-feng   

  1. School of Life Science, East China Normal University, Shanghai 200062, China
  • Received:2009-04-01 Revised:2009-08-01 Online:2010-07-25 Published:2010-07-25
  • Contact: ZHANG Hong-feng

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摘要： 灰树花（Grifola fondose）高浓缩精粉经热水提取，95%乙醇沉淀，获得水溶性灰树花粗多糖GFP.GFP依次经DEAE-cellulose阴离子交换树脂及Sephadex G-100葡聚糖凝胶分离纯化得到GFP1-F，GFP1-M及GFP1-L 3种不含蛋白质的葡聚糖纯品，其分子量依次为1.09×10^5，1.93×10^4和2.76×10^3 Da.采用吡啶-氯磺酸法对其进行硫酸酯化修饰，硫酸酯化衍生物GFP1-FS，GFP1-MS及GFP1-LS的红外光谱分析表明，3个样品均在1 236.90 cm^-1和811.81 cm^-1有硫酸酯键的特征吸收峰，13C-NMR证明C-6上的羟基被酯化.并且GFP1-FS的硫酸酯化程度最高，其取代度DS为1.07；GFP1-MS与GFP1-LS的硫酸酯化程度相当，DS分别为0.66和0.61.划痕法实验结果表明，经1 000 μg/mL 的GFP1-FS，GFP1-MS及GFP1-LS处理24 h后向划痕区迁移的细胞数明显减少，分别为对照组的73.33%，34.17%和67.21%，均具有抑制人微血管内皮细胞（HMEC）迁移的活性，其中GFP1-MS的效果最为显著，这可能与GFP1-MS所具有的复杂分支结构有关.

关键词: 灰树花, 葡聚糖, 硫酸酯化衍生物, 人微血管内皮细胞, 迁移, 灰树花, 葡聚糖, 硫酸酯化衍生物, 人微血管内皮细胞, 迁移

Abstract: Polysaccharides from Grifola frondosa (GFP) were extracted by hot water and precipitated by 95% EtOH. GFP1-F，GFP1-M and GFP1-L were further purified by DEAE-cellulose and Sephadex G-100 subsequently from GFP. GFP1-F，GFP1-M and GFP1-L were glucan with molecular weight 1.09×10^5, 1.93×10^4 and 2.76×10^3 Da. GFP1-FS, GFP1-MS and GFP1-LS were polysaccharide sulfates obtained from GFP1-F，GFP1-M and GFP1-L with chlorosulfonic acidpyridine. The IR spectrum of GFP1-FS，GFP1-MS and GFP1-LS showed the characteristic absorptions of sulfate ester bond at 1 230 cm^-1and 810 cm^-1 . The 13C-NMR results indicated the modification mainly occurred at C-6 of the polysaccharide sulfates. GFP1-MS had the greatest sulfated degree with DS 1.07, while DSs of GFP1-FS and GFP1-LS were 0.66 and 0.61 respectively. The scratching assay suggested that 73.33%，34.17% and 67.21% cells migrated to scratching area compared with control after 24hour treatment of 1 000 μg/mL GFP1-FS, GFP1-MS and GFP1-LS respectively. So that all the three sulfated derivatives had inhibitory effects on HMEC migration, especially GFP1-MS had the strongest activity which may be related with its complicated branch structure.

Key words: glucan, sulfated derivative, HMEC, migration, Grifola frondosa, glucan, sulfated derivative, HMEC, migration

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