杏鲍菇多糖的提取及其对RAW264.7细胞吞噬活性·NO释放的影响

Extraction of Pleurotus eryngii Polysaccharides and Its Effects on the Macrophages Activity of RAW264.7 Cell and the Release of NO

[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] With the physicochemical property of Pleurotus eryngii polysaccharides as the comprehensive evaluation index,polysaccharides were extracted from P. eryngii; and its effects on the macrophages activity of RAW264. 7 cell and the release of NO were researched. [Method] Polysaccharides were extracted from P. eryngii by the methods of water decoction extraction（ A）,ultrasonic extraction（ B）,90 ℃ hot water extraction（ C）,70 ℃ hot water extraction（ D） and 50 ℃ hot water extraction（ E）. Contents of total glucose,acid glucose and protein were detected by phenol-sulfuric acid method,m-hydroxydiphenyl method and lowry method. Glucose component and molecular weight distribution were detected by PMP precolumn derivatization method and HPGPC method. Phagocytic ability and NO release were detected by neutral red method and Griess kit method. [Result] Polysaccharides and total glucose contents were the maximum in 50 ℃ hot water extraction. Polysaccharides and protein contents were the maximum in ultrasonic extraction. Acid sugar content in each group was relatively low. Monosaccharide type was relatively less in polysaccharides,and molecular weight was relatively low in water decoction extraction.90 ℃ hot water extraction was the maximum extraction method. 25- 100 μg / m L significantly promoted the phagocytic ability and NO release.[Conclusion]90 ℃ hot water extraction is the optimal extraction method for P. eryngii polysaccharides. A given concentration of P. eryngii polysaccharides enhances the phagocytic activity of RAW264. 7 cells,and promotes the NO release,indicating that P. eryngii polysaccharides have certain immunocompetence.