云芝多糖对小鼠脾细胞的免疫增强作用研究

Activation of spleen cells by Coriolus Versicolor polysaccharide

目的研究云芝多糖对小鼠脾细胞的免疫增强作用。方法水提醇沉法制备云芝粗多糖,采用不同浓度的乙醇对所得云芝粗多糖进行梯度醇沉,并计算各级产物的总多糖含量及收率。将小鼠脾细胞分别与云芝粗多糖及分级醇沉各级样品共同孵育,采用CCK-8法检测各样品对脾细胞增殖、对Con A或LPS诱导的脾细胞增殖的影响,采用酶联免疫法检测各级产物对NO释放的影响。结果分级醇沉各级产物多糖含量差异较小,但收率差别较大,其中30%浓度醇沉多糖收率最高,40%浓度醇沉多糖收率最低。云芝多糖分级醇沉各级样本对小鼠脾细胞增殖均有一定的促进作用,综合评价,优选出70%、80%醇沉样本较好,且作用效果明显优于粗多糖。总多糖及70%、80%醇沉多糖样品分别在0.50~1.25、0.50~0.75、0.50~1.00 mg·m L-1内药理活性与剂量呈正相关,且对Con A或LPS诱导的脾细胞增殖及NO释放有明显的协同作用(P<0.05)。结论云芝粗多糖及分级醇沉各级产物对体外小鼠脾细胞具有免疫增强作用,本研究为云芝多糖的开发、利用研究提供了一定理论依据。

Objective To explore the immune enhancing effect of Coriolus Versicolor polysaccharide on mouse spleen cells. Methods Coriolus Versicolor polysaccharide was prepared by water extraction and alcohol precipitation method, and different concentrations of ethanol were used on the versicolor alcohol sink coarse polysaccharide by gradient. At the same time, we calculated the levels of total product yield and polysaccharide content. The spleen cells of mice with Coriolus Versicolor polysaccharides and graded ethanol precipitation samples were incubated. CCK-8 method was used to detect the effects of various samples on the proliferation of spleen cells, and the proliferation of spleen cells induced by Con A or LPS. Enzyme-linked immunoassay detection was used to evaluate the impact on the NO release at all levels. Results The difference in the content of polysaccharide at different levels of alcohol precipitation was small, but the yield varied： the yield of 30% concentration alcohol precipitation was the highest, while the yield of 40% concentration alcohol precipitation was the lowest. Coriolus Versicolor polysaccharide graded ethanol precipitation samples at all levels promoted the proliferation of mouse spleen cells. Comprehensively evaluation showed the 80% and 70% alcohol precipitation samples were optimal, whose effect was better than that of crude polysaccharide. The pharmacological activity and the dosage was positively correlated in total polysaccharides and 70%, 80% alcohol sink polysaccharide samples at 0.50-1.25, 0.50-0.75, and 0.50-1.00 mg·m L^-1 concentration range, and had obvious synergistic effect（P 0.05） on Con A or LPS induced splenocyte proliferation and NO release. Conclusion All levels of Coriolus Versicolor polysaccharides and grade alcohol sink enhance the immunity of mouse spleen cells in vitro.