Structure Characterization and Anti-inflammatory Activity of Polysaccharide ABD from Agaricus blazei Murill
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Abstract:
The structure and anti-inflammatory activity of the water-soluble polysaccharide ABD extracted from the fruit body of Agaricus blazei Murill 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 purifing 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 2.058×103 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.
