黑木耳多糖提取工艺优化及其对小鼠巨噬细胞功能的影响

采用单因素试验及正交试验，考察液料比、搅拌转速、提取时间、提取温度和原料颗粒大小（目数）对黑木耳多糖提取得率的影响，并对黑木耳多糖提取工艺进行优化。通过灌胃法给药，采用小鼠碳廓清实验和小鼠腹腔巨噬细胞吞噬荧光标记微球实验测定黑木耳多糖对小鼠巨噬细胞功能的影响。结果表明：影响黑木耳多糖提取得率的因素排序为，原料颗粒大小＞提取时间＞搅拌转速＞液料比。优化得到的黑木耳多糖提取最佳工艺条件为原料颗粒大小80 目、液料比30∶1（mL/g）、搅拌转速200 r/min、提取时间70 min、提取温度100 ℃（沸水浴加热），此条件下黑木耳多糖提取得率可达8.46%。同时，黑木耳多糖对小鼠体质量、胸腺指数和脾脏指数无显著影响，但能明显提高小鼠单核-巨噬细胞的吞噬能力，说明黑木耳多糖具有提高动物非特异性免疫能力的作用。

Optimization of Extraction Conditions of Crude Polysaccharide from Auricularia auricular and Its Effects on the Functions of Mouse Peritoneal Macrophages

In this study, the extraction conditions of water-soluble polysaccharide from dried fruiting bodies of Auricularia
auricular were investigated by single-factor and orthogonal array design methods. The influence of liquid-to-solid ratio,
stirring speed, extraction time, extraction temperature and the particle size of the dried material on extraction efficiency
was analyzed. The optimal extraction conditions were established based on the extraction yield of polysaccharide. Results
showed that the most important factor was the particle size of the dried material, followed by extraction time, stirring speed
and liquid-to-solid ratio. The optimal parameters determined in this study were as follows: the particle size, 80 mesh; liquidto-
solid ratio, 30:1(mL/g); stirring speed, 200 r/min, extraction temperature, 100 ℃; and extraction time, 70 min. Under
these conditions, the polysaccharide yield from A. auricular was 8.46%. The effects of the crude polysaccharide extracted
from A. auricular on the immunologic functions of mouse macrophages were also investigated through carbon clearance
test and peritoneal macrophage phagocytosis function test using fluorescence labeled microspheres. Additionally, the effects
of the crude polysaccharide on immune organs were also examined. Results indicated that while the crude polysaccharide
had no significant influences on the body weight, thymus index or spleen index of the animals, it significantly improved the
phagocytosis of mononuclear macrophage. In conclusion, the crude polysaccharide extracted from A. auricular can enhance
animal nonspecific immunity.