茯苓多糖硫酸酯的制备与鉴定及其增强小鼠NK细胞活性

Preparation and Characterization of Pachyman Sulfonate and its Enhancing Effect on the Killing Activity of NK Cells

以氯磺酸为磺化试剂、吡啶为溶剂半合成制备茯苓多糖硫酸酯(Pachyman sulfate,PS),对比分析磺化前后产物的特征光谱变化;以自然杀伤细胞-NK细胞体外杀伤小鼠淋巴瘤细胞Yac-1的杀伤率衡量NK细胞活性,考察PS增强NK细胞自然杀伤活性。结果表明,PS重均分子量为1.02×104Da;紫外光谱分析表明PS在260 nm处可见明显吸收峰;红外光谱分析表明,PS新出现的在1250 cm-1和810 cm-1处的吸收峰为硫酸酯化的特征峰;13C-NMR谱表明PS单糖残基上3个自由羟基均发生取代反应;PS显著提高NK细胞杀伤Yac-1细胞的活性(P<0.001),且试验范围内杀伤率随PS剂量的增加而提高。

Pachyman sulfonate （PS） （mol. wt. 1.02 × 10^4 Da） was synthesized from pachyman （dissolved in pyridine） using chlorosulfonic acid as the sulfonating reagent and the spectral characteristics before and after sulfonation were compared. A distinct absorption peak was observed at 260 nm in the UV-VIS spectrum of PS, and the FT-IR spectrum contained absorption peaks typical of sulfate-substituted glucoside hydroxyl groups at 1 250 cm-1 and 810 cm-1 . The ~3C-NMR spectrum of PS indicated complete sulfonation at C-6, C- 2 and C-4 of the glucosidc radical. PS significantly （P〈0. 001） enhanced thc killing activity of NK cells towards Yac-1 cells in vitro in a dose-dependent manner.