关黄柏总多糖的分离纯化及其对小鼠脾细胞增殖的影响

目的:分离纯化关黄柏总多糖,研究纯化组分对小鼠脾淋巴细胞增殖的影响。方法:采用DEAE Cellulose 52,DEAE-Sepharose F.F,Sephacryl S-400等载体对关黄柏总多糖进行分离纯化,高效凝胶过滤色谱法鉴定纯度并测定分子量,PMP柱前衍生化测定单糖组成;采用MTT法测定各多糖组分对Con A诱导的小鼠脾淋巴细胞增殖的作用。结果:关黄柏总多糖占药材的4.3%,从关黄柏总多糖中分离纯化得到3种多糖组分G1,G2,G3,分别占总多糖的6%,9%,10%,经HPLC鉴定证明为均一多糖,均主要含阿拉伯糖、鼠李糖、半乳糖和半乳糖醛酸,其摩尔比分别是G1(1∶1.06∶1.04∶1.14),G2(1∶1.27∶1.10∶1.09),G3(1∶1.23∶0.99∶1.55),3种均一多糖对Con A诱导的T淋巴细胞增殖有显著的抑制作用。结论:3种均一关黄柏总多糖主要由阿拉伯糖、鼠李糖、半乳糖和半乳糖醛酸组成,具有一定显著的抑制细胞免疫的作用,为进一步研究提供了实验基础。

Separation and Purification of Polysaccharides from Phellodendri Amurensis Cortex and Effect on Proliferation Function of Mice Spleen Lymphocyte

Objective: To separate and purificate homogeneous polysaccharides from Phellodendri Amurensis Cortex and study the effect on proliferation function of mice spleen lymphocyte transformation. Method: The DEAE Cellulose 52, DEAE-Sepharose F.F and Sephacryl S-400 columns were used to separate and purificate homogeneous polysaccharides and the monosaccharide composition and structure were preliminarily identificated by periodate oxidation, smith degradation, methylation, IR, HPLC and GC-MS, and their effects on proliferation function of mice spleen lymphocyte transformation were determinated by MTT. Result: The yield of crude polysaccharide was 4%. Three homogeneous polysaccharides were separated and purificated (G1-G3), the yields of purified polysaccharides were 6%, 9%, and 10%, espectively, which were identified as homogeneous polysaccharides by HPLC. They mainly contained arabinose, rhamnose, galactose and galacturonic acid, the mole ratios were 1: 1.06: 1.04: 1.14 (G1), 1: 1.27: 1.10: 1.09 (G2) and 1: 1.23: 0.99: 1.55 (G3), and the homogeneous polysaccharides were able to inhibit the proliferation of lymphocytes induced by ConA. Conclusion: The homogeneous polysaccharides contained arabinose, rhamnose, galactose and galacturonic acid, and showed inhibit effect of cellular immunity, which established the experimental basis for the further study of the polysaccharides.