Proteomic and Metabolomic Analyses Reveal Contrasting Anti-Inflammatory Effects of an Extract of Mucor Racemosus Secondary Metabolites Compared to Dexamethasone.
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.
Department of Analytical Chemistry, University of Vienna, Währinger Straße 38, 1090, Vienna, Austria.
Samuel M
SM
Meier
Department of Analytical Chemistry, University of Vienna, Währinger Straße 38, 1090, Vienna, Austria.
Besnik
B
Muqaku
Syntrion GmbH, Robert-Bosch-Straße 6, 75365, Calw, Germany.
Ronald
R
Ullmann
Department of Analytical Chemistry, University of Vienna, Währinger Straße 38, 1090, Vienna, Austria.
Andrea
A
Bileck
Department of Analytical Chemistry, University of Vienna, Währinger Straße 38, 1090, Vienna, Austria.
Dominique
D
Kreutz
Department of Analytical Chemistry, University of Vienna, Währinger Straße 38, 1090, Vienna, Austria.
Johanna C
JC
Mader
Institute of Cancer Research, Medical University of Vienna, Borschkegasse 8a, 1090, Vienna, Austria.
Siegfried
S
Knasmüller
Department of Analytical Chemistry, University of Vienna, Währinger Straße 38, 1090, Vienna, Austria.
Christopher
C
Gerner
D000595:Amino Acid Sequence; D000893:Anti-Inflammatory Agents / Q000302:isolation & purification / Q000494:pharmacology*; D001688:Biological Products / Q000302:isolation & purification / Q000494:pharmacology*; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D016207:Cytokines / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology*; D015180:Electrophoresis, Gel, Two-Dimensional; D056945:Hep G2 Cells; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism; D055432:Metabolomics / Q000379:methods*; D008969:Molecular Sequence Data; D009089:Mucor / Q000737:chemistry* / Q000378:metabolism; D009152:Mutagenicity Tests; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics; D053719:Tandem Mass Spectrometry
D000893:Anti-Inflammatory Agents; D001688:Biological Products; D016207:Cytokines; D018836:Inflammation Mediators; D020543:Proteome; D003907:Dexamethasone