关黄柏总多糖的分离纯化及其对小鼠脾细胞增殖的影响

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

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。

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 polysaceharides and the monosaeeharide composition and structure were preliminarily identifieated 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 polysaecharides contained arabinose, rhamnose, galaetose and galaeturonic acid, and showed inhibit effect of cellular immunity, which established the experimental basis for the further study of the polysaccharides.