A Polysaccharide Purified from Morchella conica Pers. Prevents Oxidative Stress Induced by H₂O₂ in Human Embryonic Kidney (HEK) 293T Cells

Int J Mol Sci. 2018 Dec 13;19(12):4027. doi: 10.3390/ijms19124027.

Abstract

Morchella conica Pers. (M. conica) has been used both as a medical and edible mushroom and possesses antimicrobial properties and antioxidant activities. However, the antioxidant properties of polysaccharides purified from M. conica have not been studied. The aim of this study was to investigate the in vitro antioxidant properties of a polysaccharide NMCP-2 (neutral M. conica polysaccharides-2) purified from M. conica, as determined by radical scavenging assay and H₂O₂-induced oxidative stress in HEK 293T cells. Results showed that NMCP-2 with an average molecular weight of 48.3 kDa possessed a much stronger chelating ability on ferrous ions and a higher ability to scavenge radical scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH) than the other purified fraction of NMCP-1 from M. conica. Moreover, 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetra-zolium bromide (MTT) assay showed that NMCP-2 dose-dependently preserved cell viability of H₂O₂-induced cells. The NMCP-2 pretreated group reduced the generation of reactive oxygen species (ROS) content and increased the mitochondria membrane potential (MMP) levels. In addition, Hoechst 33342 staining revealed cells treated with NMCP-2 declined nuclear condensation. Ultrastructural observation revealed that NMCP-2 pretreatment alleviated the ruptured mitochondria when exposed to H₂O₂. Furthermore, western blot analysis showed that NMCP-2 prevented significant downregulation of the protein expression of Bax, cleaved caspases 3, and upregulated Bcl-2 levels. These results suggest the protective effects of NMCP-2 against H₂O₂-induced injury in HEK 293T cells. NMCP-2 could be used as a natural antioxidant of functional foods and natural drugs.

Keywords: Morchella conica; oxidative stress; polysaccharides.

MeSH terms

  • Agaricales / metabolism*
  • Antioxidants / isolation & purification
  • Antioxidants / pharmacology*
  • Caspase 3 / metabolism
  • Cell Survival
  • Dose-Response Relationship, Drug
  • Fungal Polysaccharides / isolation & purification
  • Fungal Polysaccharides / pharmacology*
  • Gene Expression Regulation / drug effects
  • HEK293 Cells
  • Humans
  • Hydrogen Peroxide / adverse effects*
  • Membrane Potential, Mitochondrial / drug effects
  • Molecular Weight
  • Oxidative Stress / drug effects
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Reactive Oxygen Species / metabolism
  • bcl-2-Associated X Protein / metabolism

Substances

  • Antioxidants
  • BAX protein, human
  • BCL2 protein, human
  • Fungal Polysaccharides
  • Proto-Oncogene Proteins c-bcl-2
  • Reactive Oxygen Species
  • bcl-2-Associated X Protein
  • Hydrogen Peroxide
  • CASP3 protein, human
  • Caspase 3