论文部分内容阅读
目的:从蕨菜中分离纯化多糖成分,并进行相对分子质量、单糖组成和免疫调节活性研究。方法:采用热水浸提法提取蕨菜粗多糖,再经脱蛋白、DEAE Sepharose Fast Flow和Sepharose 4B凝胶柱层析得到3种均一多糖组分PLP1、PLP2和PLP3;采用高效液相凝胶渗透色谱法测定均一多糖相对分子质量;采用气相色谱分析均一多糖的单糖组分,采用MTT法和Griess法检测多糖对RAW264.7细胞生长和释放NO能力的影响。结果:PLP1、PLP2和PLP3的相对分子质量分别为6.62×10~4、4.34×10~5、1.88×10~5;PLP1含有鼠李糖、阿拉伯糖、岩藻糖、木糖、甘露糖、葡萄糖和半乳糖,PLP2含有鼠李糖、阿拉伯糖、岩藻糖、木糖、甘露糖和半乳糖,PLP3含有鼠李糖、阿拉伯糖、岩藻糖、木糖、甘露糖和半乳糖。3种多糖均可显著促进RAW264.7细胞的增殖,均具有显著增强RAW264.7细胞释放NO的能力,并呈一定的剂量效应关系。结论:PLP1、PLP2和PLP3 3个组分均为新的天然来源的均一杂多糖。3种多糖体外均能激活巨噬细胞,呈现显著的免疫调节活性。
Objective: To isolate and purify polysaccharides from Bracken, and to study the relative molecular mass, monosaccharide composition and immunomodulatory activity. Methods: Three kinds of polysaccharides, PLP1, PLP2 and PLP3, were obtained by hot water extraction, then deproteinized, DEAE Sepharose Fast Flow and Sepharose 4B gel column chromatography. High performance liquid gel The molecular weight of homogenous polysaccharides was determined by osmotic chromatography. Monosaccharide composition of homogenous polysaccharides was analyzed by gas chromatography. The effects of polysaccharides on the growth and the release of NO of RAW264.7 cells were detected by MTT assay and Griess assay. Results: The relative molecular masses of PLP1, PLP2 and PLP3 were 6.62 × 10 ~ 4, 4.34 × 10 ~ 5 and 1.88 × 10 ~ 5 respectively. PLP1 contained rhamnose, arabinose, fucose, xylose, mannose, PLP2 contains rhamnose, arabinose, fucose, xylose, mannose and galactose, and PLP3 contains rhamnose, arabinose, fucose, xylose, mannose and galactose. The three kinds of polysaccharides can significantly promote the proliferation of RAW264.7 cells, both significantly enhanced the ability of RAW264.7 cells to release NO, and showed a dose-response relationship. CONCLUSION: The three components of PLP1, PLP2 and PLP3 are novel homogenous heteropolysaccharides of natural origin. All 3 kinds of polysaccharides can activate macrophages in vitro, showing significant immunomodulatory activity.