姬松茸多糖ABD的结构表征及抗炎活性研究

Structure Characterization and Anti-inflammatory Activity of Polysaccharide ABD from Agaricus blazei Murill

本文研究了从姬松茸子实体中提取的水溶性多糖ABD的结构和抗炎活性。水提姬松茸多糖溶液通过sevage法除蛋白,以85%乙醇沉淀,经过DEAE-sepharose Fast Flow层析柱纯化得到多糖ABD。通过紫外全波长扫描证明其中不含蛋白质和核酸等杂质;通过刚果红试验证明其不具有三螺旋结构;通过凝胶色谱分析得出,ABD的分子量(Mw)为2.058×103 ku;通过高效液相色谱分析,其单糖组成为葡萄糖、半乳糖、甘露糖和岩藻糖(比例为83.59:5.46:0.59:1.04);通过MTT实验证明,在62.5~1000μg/mL范围内,多糖ABD对小鼠腹腔巨噬细胞(RAW264.7)没有明显的毒性;通过脂多糖诱导的小鼠体外炎症模型,测定了肿瘤坏死因子(TNF-α)和一氧化氮(NO)两种细胞因子的分泌情况,结果证明多糖ABD对这两种细胞因子的分泌呈现抑制作用,并且呈现浓度依赖性,表现出抗炎活性。

The structure and anti-inflammatory activity of the water-soluble polysaccharide ABD extracted from the fruit body of Agaricus blazei MuriU were studied in this paper. After the proteins were removed using the Sevage method, the polysaccharide ABD was obtained by precipitating with 85% ethanol （final concentration） and purifmg using the DEAE-sepharose Fast Flow chromatography column. Analysis of UV full-wavelength spectral scans showed that the ABD polysaccharide did not contain impurities, such as proteins and nucleic acids, and did not have a triple helix structure based on the result of Congo red test. The molecular weight was determined to be 2.058×10^3 ku by gel-permeation chromatography. The monosaccharide composition of ABD was evaluated by HPLC, and ABD was found to consist of glucose （83.59%）, galactose （5.46%）, mannose （0.59%）, and fucose （1.04%）. The 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay proved that the ABD polysaccharide had no obvious toxicity to macrophages （RAW264.7） within the concentration range of 62.5-1000 μg/mL. The secretion of nitric oxide and tumor necrosis factor alpha （TNF-α） was determined using a mouse model of lipopolysaccharide-induced inflammation in vitro. The results demonstrated that polysaccharide ABD had a dose-dependent inhibitory effect on the secretion of both nitric oxide and TNF-α, and exhibited excellent anti-inflammatory activity.