Proteomic and Metabolomic Analyses Reveal Contrasting Anti-Inflammatory Effects of an Extract of Mucor Racemosus Secondary Metabolites Compared to Dexamethasone

PLoS One. 2015 Oct 23;10(10):e0140367. doi: 10.1371/journal.pone.0140367. eCollection 2015.

Abstract

Classical drug assays are often confined to single molecules and targeting single pathways. However, it is also desirable to investigate the effects of complex mixtures on complex systems such as living cells including the natural multitude of signalling pathways. Evidence based on herbal medicine has motivated us to investigate potential beneficial health effects of Mucor racemosus (M rac) extracts. Secondary metabolites of M rac were collected using a good-manufacturing process (GMP) approved production line and a validated manufacturing process, in order to obtain a stable product termed SyCircue (National Drug Code USA: 10424-102). Toxicological studies confirmed that this product does not contain mycotoxins and is non-genotoxic. Potential effects on inflammatory processes were investigated by treating stimulated cells with M rac extracts and the effects were compared to the standard anti-inflammatory drug dexamethasone on the levels of the proteome and metabolome. Using 2D-PAGE, slight anti-inflammatory effects were observed in primary white blood mononuclear cells, which were more pronounced in primary human umbilical vein endothelial cells (HUVECs). Proteome profiling based on nLC-MS/MS analysis of tryptic digests revealed inhibitory effects of M rac extracts on pro-inflammatory cytoplasmic mediators and secreted cytokines and chemokines in these endothelial cells. This finding was confirmed using targeted proteomics, here treatment of stimulated cells with M rac extracts down-regulated the secretion of IL-6, IL-8, CXCL5 and GROA significantly. Finally, the modulating effects of M rac on HUVECs were also confirmed on the level of the metabolome. Several metabolites displayed significant concentration changes upon treatment of inflammatory activated HUVECs with the M rac extract, including spermine and lysophosphatidylcholine acyl C18:0 and sphingomyelin C26:1, while the bulk of measured metabolites remained unaffected. Interestingly, the effects of M rac treatment on lipids were orthogonal to the effect of dexamethasone underlining differences in the overall mode of action.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Anti-Inflammatory Agents / isolation & purification
  • Anti-Inflammatory Agents / pharmacology*
  • Biological Products / isolation & purification
  • Biological Products / pharmacology*
  • Cells, Cultured
  • Chromatography, Liquid
  • Cytokines / metabolism
  • Dexamethasone / pharmacology*
  • Electrophoresis, Gel, Two-Dimensional
  • Hep G2 Cells
  • Human Umbilical Vein Endothelial Cells / drug effects
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Inflammation Mediators / metabolism
  • Leukocytes, Mononuclear / drug effects
  • Leukocytes, Mononuclear / metabolism
  • Metabolomics / methods*
  • Molecular Sequence Data
  • Mucor / chemistry*
  • Mucor / metabolism
  • Mutagenicity Tests
  • Proteome / metabolism
  • Proteomics / methods*
  • Salmonella typhimurium / drug effects
  • Salmonella typhimurium / genetics
  • Tandem Mass Spectrometry

Substances

  • Anti-Inflammatory Agents
  • Biological Products
  • Cytokines
  • Inflammation Mediators
  • Proteome
  • Dexamethasone

Grants and funding

Syntrion GmbH funded consumables and provided the extract of M rac. Syntrion GmbH had no role in study design, data collection and analysis and the decision to publish. R.U. is grateful to Dr. Ben Warkentin and Rie Warkentin, Dr. Warkentin GmbH, Stephanskirchen as well as Dr. Herbert Reuther, Marien Apotheke, Prien for co-funding this project.