胀果甘草酸性多糖的分离纯化、结构分析及免疫活性测定

Isolation, purification, structural analysis and immune activity determination of acid polysaccharides from Glycyrrhiza inflata Bat.

中文关键词:  胀果甘草  多糖  分离纯化  结构分析  免疫活性

英文关键词:Glycyrrhiza inflata Bat.  polysaccharide  isolation and purification  structural analysis  immune activity

中文摘要:

      目的 分离纯化胀果甘草多糖, 并对得到的3种酸性多糖组分结构及免疫活性进行分析。方法 采用水提醇沉法提取胀果甘草粗多糖, 经离子交换柱层析和凝胶柱层析分离纯化, 高效凝胶渗透色谱法(high performance gel permeation chromatography, HPGPC)、红外光谱法(infrared spectroscopy, IR)、气相色谱法(gas chromatography, GC)和气相-质谱法(gas chromatography-mass spectrometry, GC-MS)分析其结构, 采用H3-胸腺嘧啶核苷掺入法(H3-TdR)测定其对小鼠淋巴细胞增殖的影响。结果 胀果甘草根及根茎经水提醇沉、离子交换柱层析和凝胶柱层析分离纯化后, 得到3种相对分子量大于2.0×106 Da的均一酸性多糖组分GiP-B1、GiP-C1和GiP-D1。结构和免疫活性的研究结果表明, 这3种多糖组分均是由阿拉伯糖(Ara)、鼠李糖(Rha)、半乳糖(Gal)、半乳糖醛酸(GalA)以不同摩尔比组成的酸性杂多糖, 多糖的主链均由1,4-GalpA和1,2-Rhap残基构成, 支链由1,5-连接的Araf和1,3-连接的Galp构成, 支链分支点位于Rhap残基的O-4位。3种多糖在体外对小鼠脾细胞增殖均有促进作用, 与空白对照组均有显著性差异(P<0.05), 其中糖醛酸含量最高的GiP-D1促脾细胞增殖作用最高。结论 从胀果甘草中分离纯化的3种酸性多糖为均一组分, 具有一定的免疫增强活性。

英文摘要:

      Objective To isolate and purify polysaccharide fractions from Glycyrrhiza inflata Bat. and analyze their basic chemical structures and immune activities. Methods Crude polysaccharides were extracted using water extracting-alcohol precipitating method, and sequentially purified by ion-exchange and gel column chromatography. Their basic chemical structures were analyzed the methods, including HPGPC (high performance gel permeation chromatography), IR(infrared spectroscopy), GC(gas chromatography) and GC-MS(gas chromatography-mass spectrometry). Their immune activities were evaluated by the H3-TdR assay. Results Three acid homogeneous polysaccharides (GiP-B1, GiP-C1, GiP-D1) were purified by ion-exchange and gel column chromatography. The molecular weights of 3 homogeneous polysaccharides were over 2.0×106 Da. They were composed of arabinose, rhamnose, galactose and galacturonic acid with different molar ratio. They might have the similar structure characteristics: the backbone was mainly made up of 1,4-linked-α-GalpA and 1,2-linked-α-Rhap residues, and the side chains were composed of 1,5-linked-α-Araf residues and 1,3-linked-α-Galp residues, mainly linked at the O-4 of α-Rhap. The results of H3-TdR assay showed that certain concentrations of 3 homogeneous polysaccharides could promote the proliferation of lymphocyte and there were significant differences (P<0.05) compared with the blank control. Conclusion GiP-B1, GiP-C1 and GiP-D1 extracted from Glycyrrhiza inflata Bat. are homogeneous polysaccharides and they have effects on enhancing immune responses.