花脸香蘑菌丝体多糖提取条件优化及其体外免疫活性

Extraction and Immune Modulatory Activities in Vitro of Polysaccharides from Lepista sordida Mycelia

为探讨花脸香蘑菌丝体多糖提取工艺及其免疫活性,以花脸香蘑菌丝体为原料,在单因素试验的基础上采用正交试验优化多糖提取条件;将不同质量浓度的花脸香蘑菌丝体多糖溶液作用于小鼠巨噬细胞Ana-1,通过对其细胞存活率、细胞因子TNF-α和IL-6分泌量、中性红吞噬能力以及膜表面蛋白Toll样受体(Toll-like receptor,TLR)TLR2和TLR4表达量能力的测定,研究花脸香蘑菌丝体多糖的免疫活性。结果表明,花脸香蘑菌丝体多糖最佳提取条件为浸提温度65℃、浸提时间2.5 h、液料比40∶1(m L/g),提取率最高为15.88%;在0.05~1.6 mg/m L的有效剂量范围内,花脸香蘑菌丝体多糖能够不同程度地促进小鼠巨噬细胞Ana-1的增殖,提高细胞因子TNF-α和IL-6分泌量,增强中性红吞噬能力、膜表面蛋白TLR2和TLR4表达量能力,其中以1.6 mg/m L效果最佳。表明花脸香蘑菌丝体多糖具有优良的免疫活性。

This study aimed to improve the extraction yield and evaluate the immune modulatory activity of polysaccharides of Lepista sordida mycelia.Optimization of the extraction conditions was carried out using one-factor-at-a-time and orthogonal array design methods.The immune modulatory effect of the polysaccharides extracted was evaluated by measuring cell survival,TNF-αand IL-6secretion,the percentage of neutral red phagocytosis and the expression levels of Toll-like receptor(TLR)TLR2and TLR4when mouse Ana-1macrophages were exposed to different concentrations of the polysaccharides.The optimal extraction conditions were determined as follows:extraction temperature,65℃;extraction time,2.5h;solvent-to-solid ratio,40:1(mL/g),leading to an extraction yield of15.88%.In the concentration range of0.05-1.6mg/mL,the polysaccharides could promote the proliferation of macrophages,TLR2and TLR4expression,the phagocytosis of neutral red and TNF-αand IL-6production;the most effective concentration was1.6mg/mL.These data illustrated that the polysaccharides possessed good immune modulatory activity.