摘要
目的：分离纯化胀果甘草多糖,并研究其结构及免疫活性。方法：采用水提醇沉法提取胀果甘草粗多糖,经离子交换柱色谱和凝胶柱色谱分离纯化,高效凝胶渗透色谱（HPGPC）法、红外光谱（IR）法、气相-质谱（GC-MS）法和核磁共振氢谱（-1H-NMR）法分析其结构,噻唑蓝（MTT）法测定其对小鼠淋巴细胞和巨噬细胞增殖的影响。结果：胀果甘草根及根茎经水提醇沉,离子交换柱色谱和凝胶柱色谱分离纯化,得到平均相对分子质量为2.1×10^4Da的均一多糖组分GiP-3。对该多糖的结构和免疫活性的研究结果表明,GiP-3是由阿拉伯糖（Ara）,鼠李糖（Rha）,半乳糖（Gal）以1∶0.12∶18组成的中性阿拉伯半乳聚糖,其结构的主链由→3）α-Galp（1→残基组成,→5）α-Araf（1→支链和少量→2,4）α-Rhap（1→支链位于α-Galp残基的O-6位上。GiP-3在体外对未经诱导和经刀豆蛋白（ConA）或脂多糖（LPS）诱导的小鼠脾细胞增殖均有促进作用,对RAW 264.7巨噬细胞的增殖也有一定的促进作用,与空白对照组均有显著性差异（P〈0.05）。结论：GiP-3为均一多糖,具有免疫增强活性。
Objective： To isolate and purify a polysaccharide fraction from Glycyrrhiza inflate,in order to analyze its basic chemical structure and immune activity.Method： Crude polysaccharide was extracted by the water extracting-alcohol precipitating method,and purified by DEAE cellulose-52 ion-exchange and Sepharose CL-6B gel column chromatography.Neutral homogeneous polysaccharide GiP-3 was purified from the water eluate fraction.HPGPC,IR,GC-MS and -1H-NMR were applied to analyze the basic chemical structure.3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide（MTT） assay was applied to evaluate the immune activity.Result： The molecular weight of GiP-3 was 2.1 × 10^4 Da.It was composed of arabinose,rhamnose and galactose,with the molar ratio of 1 ∶ 0.12 ∶ 18.The backbone of GiP-3 was mainly made up of 1,3-linked-α-Galp residues,and the side chains were composed of 1,5-linked-α-Araf residues and 1,2,4-linked-α-Rhap residues,and mainly linked at O-6 of α-Galp residue.The results of MTT assay showed that certain concentrations of GiP-3 could promote the proliferation of lymphocyte and RAW264.7 macrophage,with significant differences（P〈0.05）.Conclusion：GiP-3 is a homogeneous polysaccharide extracted from G.inflate,with an effect in enhancing immune responses.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2018年第4期66-71,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81360626)
关键词
胀果甘草
多糖
分离纯化
结构分析
免疫活性
Glycyrrhiza inflate polysaccharide
isolation and purification
structural analysis
immune activity
