Thymoquinone increases the expression of neuroprotective proteins while decreasing the expression of pro-inflammatory cytokines and the gene expression NFκB pathway signaling targets in LPS/IFNγ -activated BV-2 microglia cells
Graphical abstract
Section snippets
List of abbreviations
3-MST 3 mercaptopyruvate sulfurtransferase AD Alzheimer's disease ANOVA analysis of variance ATP adenosine triphosphate BVR-A biliverdin reductase A C3 complement component 3 CCL5 chemokine (CC) motif ligand 5 CFB complement factor B CNS central nervous system CXCL3 chemokine (CC motif) ligand 3 Grx glutaredoxin GSH glutathione IFNγ interferon gamma IκB inhibitor of kappa B IL interleukin LC-MS/MS liquid chromatography with tandem mass spectrometry LONM lon protease, mitochondrial homolog LPS lipopolysaccharide
Materials
High glucose Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 4 mM GlutaMAX™, penicillin-streptomycin (10,000 U/ml), interferon gamma recombinant mouse protein (IFNγ), and trypsin/EDTA (0.25%) with phenol red were purchased from Thermo Fisher Scientific. Heat-inactivated fetal bovine serum (FBS) was purchased from Atlanta Biologicals. TQ (99% purity), lipopolysaccharides from Escherichia coli (LPS), and the reagents and Microcon-30 kDa centrifugal filter units used in the sample
Results
Comparative quantitative proteomic analysis of LPS/IFNγ-activated BV-2 cells with and without TQ treatment revealed 35 differentially expressed proteins (>95% identification confidence). Amongst these differentially expressed proteins, TQ treatment (12.5 μM for 24 h) of the LPS/IFNγ-activated microglia compared to the untreated, activated microglia resulted in the increased expression of 4 neuroprotective proteins: glutaredoxin-3 (21 fold), biliverdin reductase A (15 fold), 3-mercaptopyruvate
Discussion
Neuroinflammation has been increasingly implicated in the onset and progression of multiple neurodegenerative diseases, such as Alzheimer's disease (AD), Parkinson's disease (PD) and Multiple Sclerosis (MS), despite their differing pathologies (Liu and Hong, 2003; Block and Hong, 2005; Gao and Hong, 2008; Chen et al., 2016; Kempuraj et al., 2016). Neuroinflammation is an innate, and initially, protective response mechanism in the brain, facilitated mostly by microglia and astrocytes producing a
Conclusion
Our findings are the first to show that TQ treatment in the activated BV-2 microglial cells increased the expression of antioxidant and neuroprotective proteins, biliverdin reductase-A, 3-mercaptopyruvate sulfurtransferase, glutaredoxin-3, and mitochondrial lon protease. TQ also reduced the expression of several inflammatory cytokines in the LPS/IFNγ activated BV-2 microglial cells. Furthermore, our studies showed TQ modulated the expression of genes involved in the NFκB signaling pathway,
Acknowledgments
This research was supported by NIH-National Institute on Minority Health and Health Disparity Grants G12 MD007582 and P20 MD 006738.
Conflict of interest
None declared.
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