Purification, characterization and immunostimulatory effects of polysaccharides from Anemarrhena asphodeloides rhizomes

Int J Biol Macromol. 2021 Mar 1:172:550-559. doi: 10.1016/j.ijbiomac.2021.01.088. Epub 2021 Jan 16.

Abstract

The crude polysaccharide was extracted from A. asphodeloides rhizomes and further purified to produce two fractions F1 (50.0%) and F2 (19.6%). The chemical constitutions of the polysaccharides were neutral sugars (51.4%-89.7%), uronic acids (1.0%-30.2%) and sulfate esters (3.4%-8.1%), with various ratios of monosaccharides including rhamnose (1.4%-6.1%), arabinose (7.1%-21.2%), xylose (0.2%-4.8%), mannose (39.9%-79.0%), glucose (6.0%-11.1%) and galactose (2.6%-22.0%). The molecular properties of the polysaccharides were investigated by the HPSEC-UV-MALLS-RI system, revealing the Mw 130.0 × 103-576.5 × 103 g/moL, Rg 87.6-382.6 nm and SVg 0.3-54.3 cm3/g. The polysaccharides stimulated RAW264.7 cells to produce considerable amounts of NO and up-regulate the expression of TNF-α, IL-1 and COX-2 genes. Polysaccharides exhibited the growth inhibitory effects on cancer cells lines of AGS, MKN-28 and MKN-45, in which F2 fraction exhibited prominent bioactivities. The AGS cells treated with F2 experienced condensed cytoplasm, shrinkage of nucleus and chromatin marginalization with the highest number of cells at early-stage apoptosis reaching 54.6%. The inhibitory effect of F2 polysaccharide on AGS cells was through MAPKs and STAT3 signaling pathways. The backbone of the F2 was mainly linked by (1 → 4)-linked mannopyranosyl and (1 → 3)-linked galactopyranosyl. Taken together, the polysaccharide from A. asphodeloides rhizomes could be utilized as medicinal, pharmacological and functional food ingredients.

Keywords: Characterization; Immunostimulatory; Polysaccharides.

MeSH terms

  • Anemarrhena / chemistry*
  • Animals
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Carbohydrate Sequence
  • Cell Line, Tumor
  • Chromatin / chemistry
  • Chromatin / drug effects
  • Chromatin / immunology
  • Cyclooxygenase 2 / genetics
  • Cyclooxygenase 2 / immunology
  • Cytoplasm / drug effects
  • Cytoplasm / immunology
  • Cytoplasm / pathology
  • Gene Expression Regulation / drug effects*
  • Immunologic Factors / chemistry
  • Immunologic Factors / isolation & purification
  • Immunologic Factors / pharmacology*
  • Interleukin-1 / genetics
  • Interleukin-1 / immunology
  • Mice
  • Mitogen-Activated Protein Kinases / genetics
  • Mitogen-Activated Protein Kinases / immunology
  • Monosaccharides / chemistry
  • Monosaccharides / isolation & purification
  • Nitric Oxide / biosynthesis
  • Polysaccharides / chemistry
  • Polysaccharides / isolation & purification
  • Polysaccharides / pharmacology*
  • RAW 264.7 Cells
  • Rhizome / chemistry*
  • STAT3 Transcription Factor / genetics
  • STAT3 Transcription Factor / immunology
  • Signal Transduction
  • Tumor Necrosis Factor-alpha / genetics
  • Tumor Necrosis Factor-alpha / immunology
  • Uronic Acids / chemistry
  • Uronic Acids / isolation & purification

Substances

  • Chromatin
  • Immunologic Factors
  • Interleukin-1
  • Monosaccharides
  • Polysaccharides
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • Tumor Necrosis Factor-alpha
  • Uronic Acids
  • Nitric Oxide
  • Ptgs2 protein, mouse
  • Cyclooxygenase 2
  • Mitogen-Activated Protein Kinases