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目的从冬凌草Rabdosia rubescens中提取分离获得多糖组分RPPSIIa,并分析其结构性质和探讨免疫活性,为冬凌草临床应用与进一步开发利用提供参考。方法主要采用水提醇沉透析得冬凌草多糖,经分级醇沉,Sephdex G-100凝胶色谱柱纯化得RPPSIIa,应用HPLC、IR、LC、UV等方法对冬凌草多糖进行组成结构分析及理化性质研究,通过MTT比色法观察该组分在体外对小鼠脾淋巴细胞增殖的影响。结果冬凌草多糖RPPSIIa为均一组分,其单糖组成为鼠李糖、葡萄糖,二者比为7︰93,RPPSIIa具有典型的多糖吸收峰,其结构中存在α-型糖苷键;RPPSIIa在5～100μg/mL能有效刺激小鼠脾淋巴细胞增殖,5、50μg/mL均能有效增强刀豆蛋白A(ConA)诱导的淋巴细胞增殖反应。结论多糖RPPSIIa是由冬凌草中提取的均一葡聚糖,具有较强的免疫生物活性。
OBJECTIVE To extract and isolate RPPSIIa from Rabdosia rubescens and analyze its structure and immunological activity, providing a reference for the clinical application and further development and utilization of Rubescens. Methods The polysaccharide of Rubescens mongolicum was dialyzed by water and alcohol precipitation, purified by Sephadex G-100 gel column and fractionated by alcohol precipitation, and purified by HPLC, IR, LC, UV and so on. And physical and chemical properties of MTT colorimetric method to observe the components of mouse spleen lymphocyte proliferation in vitro. Results Rubescens polysaccharide RPPSIIa as a homogeneous component, the monosaccharide composition of rhamnose, glucose, the ratio between the two is 7: 93, RPPSIIa has a typical polysaccharide absorption peak, the structure exists α-glycosidic bond; RPPSIIa 5 ~ 100μg / mL can effectively stimulate the proliferation of splenic lymphocytes in mice, 5,50μg / mL can effectively enhance Con A induced lymphocyte proliferation. Conclusion Polysaccharide RPPSIIa is a homogeneous dextran extracted from Rubescens indica and has strong immunogenicity.