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目的:从不同发育阶段的霍山石斛组织培养物中提取分离多糖,比较多糖的理化性质和免疫活性。方法:采用水提醇沉法提取多糖,阴离子交换层析法分离多糖,分光光度法、GC-MS、IR、荧光电泳分析多糖的理化性质,ELISA法测定多糖的免疫活性。结果:霍山石斛不同发育阶段培养物中多糖含量无显著差异,经阴离子交换层析均得到5种多糖组分,且都是由葡萄糖、半乳糖和甘露糖组成,IR和荧光电泳分析显示它们的化学结构特性无显著变化;体外实验表明,霍山石斛不同发育阶段培养物多糖具有相似的促进小鼠脾细胞产生IFN-γ和TNF-α的作用。结论:组织快繁中霍山石斛不同发育阶段培养物中多糖的理化性质和免疫活性无显著差异,表现出一定的稳定性。
OBJECTIVE: To isolate polysaccharide from the tissue culture of Dendrobium huoshanense at different developmental stages and to compare the physicochemical properties and immunological activities of polysaccharides. Methods: The polysaccharides were extracted by water extraction and alcohol precipitation, the polysaccharides were separated by anion exchange chromatography, the physicochemical properties of polysaccharides were analyzed by spectrophotometry, GC-MS, IR and fluorescence electrophoresis. The immunological activity of polysaccharides was determined by ELISA. Results: There were no significant differences in the content of polysaccharides in different developmental stages of Dendrobium huoshanense. Five kinds of polysaccharides were obtained by anion exchange chromatography, and all of them were composed of glucose, galactose and mannose. IR and fluorescence electrophoresis showed that There was no significant change in the chemical structure of the cells. In vitro experiments showed that polysaccharides from different culture stages of Dendrobium huoshanense had the similar effects of promoting the production of IFN-γ and TNF-α in mouse spleen cells. CONCLUSION: There was no significant difference in the physicochemical properties and immunological activities of polysaccharides in the different developmental stages of D. huoshanense in the rapid propagation of tissues, showing some stability.