Intestinal immune system modulating polysaccharides from rhizomes of Atractylodes lancea

Hot water extract (ALR-0) of rhizomes of Atractylodes lancea DC was fractionated into MeOH-soluble fraction (ALR-1), supernatant fraction of EtOH precipitation (ALR-3+4), and crude polysaccharide fraction (ALR-5). Among these fractions. only ALR-5 showed potent stimulating activity for proliferation of bone marrow cells mediated by Peyer's patch cells. ALR-5 gave three potently active carbohydrate-rich fractions (ALR-5IIa, 5IIb, and 5IIc) by anion-exchange chromatography on DEAE-Sepharose CL-6B, and three active polysaccharides (ALR5IIa-1-1, ALR-5IIb-2-2, and ALR-5IIc-3-1) were further purified from the respective fractions. The order of activity was revealed to be ALR-5IIb-2-2 greater than or equal to ALR-5IIa-1-1 > ALR-5IIc-3-1. ALR-5IIa-1-1, 5IIb-2-2, and 5IIc-3-1 each was eluted as a single peak on HPLC and their molecular weights were estimated to be 74,000, 3,100, 16,000, respectively ALR-5IIa-1-1 consisted mainly of Am and Gal (molar ratio; 0.6:1.0) in addition to a trace amount of uronic acid whereas ALR-5IIb-2-2 and ALR-5IIc-3-1 mainly comprised Am, Gal, GlcA, and GalA (molar ratio; 0.2:1.0 : 0.2 : 0.8, and 0.5 : 1.0 : 0.7 : 1.5, respectively). Methylation analysis indicated that ALR-5IIa-1-1 consisted mainly of terminal Araf, 4- or 5-linked Ara, 3,4- or 3,5-branched Ara, and 3-linked, 4-linked, and 3,6-branched Gal. ALR-5IIb-2-2 and ALR-5IIc-3-1 were composed mainly of terminal Araf, 4- or 5-linked Ara, 4-linked Gal, 4-linked GalA, and terminal GlcA. In addition, ALR-5IIb-2-2 mainly comprised 4-linked Xyl whereas ALR-5IIc-3-1 consisted mainly of 2,4-branched Rha. Single radial gel diffusion indicated that ALR-5IIa-1-1 showed a strong reactivity with beta-glucosyl-Yariv antigen, whereas ALR-5IIb-2-2 and ALR-5IIc-3-1 did not show the reactivity with the antigen. Treatments of ALR-5IIa with NaIO4, NaClO2 and pronase did not reduce the stimulating activity for Peyer's patch cells. however combination of exo-alpha-L-arabinofuranosidase and exo-beta-D-(1-->3)-galactanase digestions of ALR-5IIa-1-1 significantly decreased its activity.