A potent sphingomyelinase inhibitor from Cordyceps mycelia contributes its cytoprotective effect against oxidative stress in macrophages

Shwu Huey Wang, Wen Bin Yang, Yin Chen Liu, Yi Hua Chiu, Chien Tsu Chen, Pai Feng Kao, Chun Mao Lin

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

A novel water-soluble polysaccharide fraction, CME-1, with a molecular mass of 27.6 kDa and containing mannose and galactose in a respective ratio of 4:6, was prepared from Cordyceps sinensis mycelia and identified by NMR and GC-MS. In the current study, we examined whether CME-1 has anti-inflammatory effects in RAW264.7 cells. The ability of CME-1 to inhibit H2O 2-induced cell death in RAW264.7 cells was assessed by using an MTT assay and annexin V/propidium iodide double staining; we found that CME-1 protected cells against H2O2-induced injury. H 2O2-induced intracellular oxidative stress and mitochondrial membrane depolarization were also diminished with CME-1 treatment. We evaluated the hydroxyl radical scavenging ability of CME-1 by using the DMPO-electron spin resonance technique, which indicated that CME-1 acts as an intracellular antioxidant in a concentration-dependent manner through a mechanism other than its scavenging activity. Activities of both neutral and acid sphingomyelinases (SMases) were assessed in vitro, and results showed that the CME-1 inhibited activities of both neutral and acid SMases in a concentration-dependent manner. CME-1 reduced H2O2 treatment-elevated C16- and C18-ceramide levels measured by LC/MS/MS in RAW264.7 cells. Results suggest that CME-1 protects RAW264.7 cells against oxidative stress through inhibition of SMase activity and reduction of C16-and C18-ceramide levels.

Original languageEnglish
Pages (from-to)471-479
Number of pages9
JournalJournal of Lipid Research
Volume52
Issue number3
DOIs
Publication statusPublished - Mar 2011

Fingerprint

Cordyceps
Sphingomyelin Phosphodiesterase
Oxidative stress
Macrophages
Ceramides
Mycelium
Scavenging
Oxidative Stress
Acids
Propidium
Annexin A5
Depolarization
Molecular mass
Cell death
Mannose
Galactose
Hydroxyl Radical
Polysaccharides
Paramagnetic resonance
Assays

Keywords

  • Ceramide
  • Cordyceps sinensis mycelia
  • Macrophage
  • Polysaccharide
  • Reactive oxygen species
  • Sphingomyelin

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Endocrinology

Cite this

A potent sphingomyelinase inhibitor from Cordyceps mycelia contributes its cytoprotective effect against oxidative stress in macrophages. / Wang, Shwu Huey; Yang, Wen Bin; Liu, Yin Chen; Chiu, Yi Hua; Chen, Chien Tsu; Kao, Pai Feng; Lin, Chun Mao.

In: Journal of Lipid Research, Vol. 52, No. 3, 03.2011, p. 471-479.

Research output: Contribution to journalArticle

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