白术多糖调节IEC-6细胞迁移活性成分的分离和表征

摘要

Bioassay-guided fractionation led to isolation of a new polysaccharide YY13008 from the roots of Atractylodes macrocephala Koidz by aqueous extraction, ethanol precipitation and purification followed by column chromatography on DEAE-cellulose and sephadex LH-20 . The chemical structure of polysaccharide YY13008 was then elucidated on the basis of acidic hydrolysis, environmental scanning electron microscope and spectroscopic data, including IR, 1D and 2D NMR analysis. The polysaccharide YY13008 was deter-mined as homogeneous, with a molecular weight of 6546, as determined by a gel permeation chromatography-multi-angle laser light scattering( GPC-MALLS) method. The polysaccharide YY13008 was found to be a fruc-tan containing almost exclusively a(2→1) -linkedβ-D-fructofuranosyl repeating units with terminalα-glucopy-ranosyl unit. The in vitro wound healing assays revealed that the treatment of polysaccharide YY13008 with di-fluoromethylornithine( DFMO) , a specific inhibitor of ornithine decarboxylase, was able to completely reverse DFMO-induced inhibition of IEC-6 migration, however, the treatment of YY13008 alone did not affect the basal rate of IEC-6 migration.%基于药理活性导向法,利用水提醇沉、DEAE纤维素柱和Sephadex LH-20葡聚糖凝胶柱等分离技术从白术根部位筛选分离出1个新的多糖组分YY13008,并利用酸水解、凝胶渗透色谱-多角度激光散射联用技术、环境扫描电子显微镜、傅里叶变换红外光谱( FTIR)、核磁共振波谱( NMR)等表征了其分子结构。表征结果表明, YY13008为线团型均一性多糖组分,绝对分子量为6546,其分子结构是由大量果糖单元以β(2→1)果糖苷键,终末端以α(1→2)葡萄糖苷键连接构成的果聚糖。体外伤口愈合细胞模型测试结果表明,白术多糖组分YY13008可使由α-二氟甲基鸟氨酸所致的IEC-6细胞迁移抑制恢复至正常水平,而其自身对IEC-6细胞迁移无明显影响。