醋柴胡多糖VBCP-3的结构和抗炎作用研究

Study on the Structure and Anti-inflammatory Effect of Vinegar Bupleurum chinese Polysaccharide VBCP-3

分离纯化醋柴胡多糖,分析其结构并探讨其对脂多糖诱导的小鼠巨噬细胞RAW264.7炎症反应的影响。用水提醇沉法提取醋柴胡粗多糖;Sevage法脱蛋白,再通过DEAE-Sepharose fast flow柱层析纯化得到醋柴胡精多糖,紫外扫描法和高效凝胶渗透色谱法测定精多糖的纯度及分子量,红外光谱法确定糖链上的官能团类型,GC-MS法测定单糖组成,MTT法测定精多糖对巨噬细胞RAW264.7存活率的影响,Griess试剂法测定精多糖对LPS诱导小鼠巨噬细胞的NO分泌量的影响。由此得到醋柴胡精多糖VBCP-3,VBCP-3为吡喃糖,不含蛋白质和核酸,分子质量为436.221 ku,其单糖组成为鼠李糖∶阿拉伯糖∶半乳糖∶葡萄糖=1.3∶24.9∶3.2∶1.00。VBCP-3对巨噬细胞RAW264.7无显著毒性。与模型组比较,VBCP-3能显著抑制NO的释放量(P<0.01)。醋柴胡多糖VBCP-3是一种单糖组成丰富的杂多糖,具有一定的抗炎活性。

To isolate and purify polysaccharide from vinegar Bupleurum chinese,analyze its structure and explore its anti-inflammatory activity in mouse RAW264.7 induced by lipopolysaccharide(LPS),the crude polysaccharide was extracted from vinegar Bupleurum chinese,precipitated with ethanol,and then the protein was removed by Sevage method.After that,the purified polysaccharide was obtained through the DEAE-Sepharose Fast Flow column chromatography.Next,the purity,molecular weight,functional group type,monosaccharide composition of the purified polysaccharide,the cell viability of the purified polysaccharide assay on RAW264.7,and the content of NO secreted by RAW264.7 were determined,respectively.The purified polysaccharide(VBCP-3)with a molecular weight of 436.221 ku consists of rhamnose,arabinose,galactose and glucose in the ratio of 1.3∶24.9∶3.2∶1.00.MTT assay showed that VBCP-3 had no obvious toxicity on RAW264.7 cells.Compared with the model group,VBCP-3 significantly inhibited the release of NO(P<0.01).VBCP-3 is a heteropolysaccharide which has obvious anti-inflammatory activity.