Separation, purification, structural analysis and immune-enhancing activity of sulfated polysaccharide isolated from sea cucumber viscera

Int J Biol Macromol. 2020 Jul 15:155:1003-1018. doi: 10.1016/j.ijbiomac.2019.11.064. Epub 2019 Nov 9.

Abstract

A novel sulfated polysaccharide (SCVP-1) was isolated from sea cucumber viscera and purified to elucidate its structure and immune-enhancing ability. SCVP-1 was found to be a homogeneous polysaccharide with a relative molecular weight of 180.8 kDa and composed of total sugars (60.2 ± 2.6%), uronic acid (15.3 ± 1.8%), proteins (6.8 ± 0.8%), and sulfate groups (18.1 ± 0.9%). SCVP-1 consisted of mannose, glucosamine, glucuronic acid, N-acetyl-galactosamine, glucose, galactose and fucose at an approximate molar ratio of 1.00:1.41:0.88:2.14:1.90:1.12:1.24. The fourier transform infrared spectra (FT-IR) and nuclear magnetic resonance (NMR) analyses showed that SCVP-1 was a kind of glycosaminoglycan. And the sulfation patterns of the fucose branches were Fuc2,4S, Fuc3,4S and Fuc0S. The surface morphology of SCVP-1 presented loose and irregular sheet structure formed by aggregation of polysaccharide molecules with spherical structure. Moreover, SCVP-1 promoted the production of nitric oxide (NO) and cytokines (IL-1β, IL-6 and TNF-α) by RAW264.7 cells as well as the expression of related genes (iNOS, IL-1β, IL-6 and TNF-α) and also enhanced their phagocytic activity through TLR4-mediated activation of the MAPKs and NF-κB signaling pathways. This study suggests that sea cucumber viscera are good sources of polysaccharides and SCVP-1 might be a novel immunomodulator.

Keywords: Immune enhancement; RAW264.7 macrophages; Sea cucumber viscera; Structure; Sulfated polysaccharide.

MeSH terms

  • Animals
  • Cell Line
  • Cytokines / metabolism
  • Immunologic Factors / chemistry
  • Immunologic Factors / isolation & purification
  • Immunologic Factors / pharmacology*
  • Macrophages / drug effects
  • Macrophages / immunology*
  • Macrophages / metabolism
  • Mice
  • NF-kappa B / metabolism
  • Polysaccharides / chemistry
  • Polysaccharides / isolation & purification*
  • Polysaccharides / pharmacology*
  • Sea Cucumbers / chemistry*
  • Sea Cucumbers / immunology
  • Sea Cucumbers / metabolism
  • Sulfates / chemistry*
  • Toll-Like Receptor 4 / metabolism
  • Viscera / chemistry*
  • Viscera / immunology
  • Viscera / metabolism

Substances

  • Cytokines
  • Immunologic Factors
  • NF-kappa B
  • Polysaccharides
  • Sulfates
  • Tlr4 protein, mouse
  • Toll-Like Receptor 4