黑松露多糖分离纯化与抗炎活性研究

以黑松露为原料,通过水提醇沉法提取黑松露多糖。提取的黑松露多糖经过DEAE Sepharose Fast Flow离子交换层析柱分离得到两个块菌多糖组分,分别为TP1和TP2。检测TP1和TP2对小鼠巨噬细胞(RAW264.7)增殖抑制作用、中性红吞噬能力影响。TP1进一步用葡聚糖凝胶层析柱纯化,得TP1-1并进行结构表征。结果表明:黑松露多糖两种组分TP1和TP2对RAW264.7细胞没有毒性作用,且都能显著增强RAW264.7细胞的中性红吞噬能力,但TP1的作用优于TP2。TP1-1的数均分子量为757910 u,其单糖组成为D-葡萄糖和D-甘露糖,比例分别为88.94%和1.19%。紫外光谱扫描结果显示块菌多糖TP1-1不含蛋白质和核酸。红外光谱分析结果表明,TP1-1具有吡喃环的结构,具有α-型糖苷键,含有甘露糖。

Isolation, Purification, and Characterization of Anti-inflammatory Activity of Truffle Polysaccharides

Using Yunnan truffle as the raw material, black truffle polysaccharides (TPs) were obtained by water extraction and alcohol precipitation. The extracted black TPs were separated by DEAE Sepharose Fast Flow ion exchange chromatography to obtain two truffle polysaccharide fractions: TP1 and TP2. The inhibitory effects of TP1 and TP2 on the proliferation of mouse macrophages (RAW264.7) and the capacities of TP1 and TP2 for the phagocytosis of neutral red pigment were investigated. TP1 was further purified by Sephadex chromatography column to yield TP1-1, which was used for structural characterization. The results showed that TP1 and TP2 had no toxic effects on RAW264.7 cells; they could significantly increase the phagocytic capacity of RAW264.7 cells, and the effect of TP1 was greater than that of TP2. The average molecular weight of TP1 was 757910 u, and its monosaccharide composition was D-glucose and D-mannose, in proportions of 88.94% and 1.19%, respectively. Ultraviolet (UV) spectra revealed that TP1-1 did not contain protein or nucleic acid. The results of infrared spectrum analysis showed that TP1-1 had a pyran ring structure and contained alpha-glycosidic bonds and mannose.