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[目的]以杏鲍菇多糖的理化性质作为综合评价指标,提取杏鲍菇多糖,并研究其对RAW264.7细胞吞噬活性的作用及对NO释放的影响。[方法]采用水煎煮提取、超声提取、90℃热水浸提、70℃热水浸提、50℃热水浸提提取杏鲍菇多糖,并采用苯酚-硫酸法、间羟基联苯法以及Lowry法测定其总糖、酸性糖和蛋白质含量;采用PMP柱前衍生化法和HPGPC法测定其组成糖和分子量分布;采用吞噬中性红法和Griess试剂盒法测定其吞噬能力及NO的释放量。[结果]试验得出,50℃热水浸提提取的多糖总糖含量最高,超声提取的多糖蛋白质含量最高,各组提取酸性糖含量相当且均较低,水煎煮提取多糖中的单糖种类最少、分子量最低。综合比较,90℃热水浸提为杏鲍菇多糖的最佳提取方法;在质量浓度为25~100μg/m L,能够极显著地促进细胞吞噬中性红的能力和NO的释放。[结论]90℃热水浸提水煎煮为杏鲍菇多糖的最佳提取方法,一定浓度的杏鲍菇多糖能够提高RAW264.7细胞的吞噬活性,并能促进NO的释放,提示杏鲍菇多糖具有一定的免疫活性。
[Objective] Based on the physicochemical properties of Pleurotus eryngii polysaccharide as a comprehensive evaluation index, the Pleurotus eryngii polysaccharide was extracted and its effect on the phagocytic activity of RAW264.7 cells and its effect on NO release were studied. [Method] The extraction of polysaccharides from Pleurotus eryngii was carried out by decoction with boiling water, ultrasonic extraction, hot water extraction at 90 ℃, hot water extraction at 70 ℃ and hot water extraction at 50 ℃. The polysaccharides were extracted by phenol - sulfuric acid method and m - And the content of total sugar, acid sugar and protein in Lowry method were determined. The compositional sugar and molecular weight distribution were determined by PMP pre-column derivatization and HPGPC method. The phagocytic capacity and NO of phagocytosis were determined by phagocytic neutral method and Griess kit Release volume. [Result] The results showed that the total sugar content of polysaccharide extracted by hot water extraction at 50 ℃ was the highest, the content of polysaccharide extracted by ultrasonic was the highest, the content of acid sugar in each group was quite low and both were lower. The smallest species, the lowest molecular weight. Taken together, hot water extraction at 90 ℃ is the best method for extracting polysaccharides from Pleurotus eryngii. When the concentration is 25-100 μg / m L, the ability of phagocytosis of neutral red and the release of nitric oxide can be remarkably promoted. [Conclusion] The optimal extraction method for polysaccharides from Pleurotus eryngii at 90 ℃ in hot water was as follows: a certain concentration of Pleurotus eryngii polysaccharide could increase the phagocytic activity of RAW264.7 cells and promote the release of NO, suggesting that Pleurotus eryngii Mushroom polysaccharide has a certain degree of immunocompetence.