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Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells
Published on: May 22, 2014
Nrf2 activator Diethyl Maleate attenuates ROS mediated NLRP3 inflammasome activation in murine microglia
Cagla Kiser1,2, Ceren Perihan Gonul1,2, Sermin Genc1,3
1Izmir Biomedicine and Genome Center, Dokuz Eylul University Health Campus, Mithatpasa St. 58/5 Balcova, 35340 Izmir, Turkey.
Abstract:
Microglia are the tissue-resident immune cells of the central nervous system. As a part of the innate immune response, NLR Family Pyrin Domain Containing Protein 3 (NLRP3) inflammasome activation leads to cleavage of caspase-1 and triggers secretion of proinflammatory cytokines and may also result in pyroptotic cell death. Inflammasome activation plays a crucial role in inflammatory conditions; aberrant activation of inflammasome contributes to the pathogenesis of neurodegenerative diseases. Diethyl Maleate (DEM) is a promising antiinflammatory chemical to alleviate inflammasome activation. In this study, NLRP3 inflammasome was activated in N9 murine microglia via 1 µg/ml LPS (Lipopolysaccharide) for 4 h and 5 mM ATP (Adenosine 5'-triphosphate) for 1 h, respectively. We demonstrated that 1 h pretreatment of DEM attenuated NLRP3 inflammasome activation in microglial cells. Besides, mitochondrial ROS decreased upon DEM pretreatment in inflammasome-induced cells. Likewise, it ameliorated pyroptotic cell death in microglia. DEM is a potent activator of Nrf2 transcription factor, the key regulator of the antioxidant response pathway. Nrf2 has been a significant target to decrease aberrant inflammasome activation through the antioxidant compounds, including DEM. Here, we have shown that DEM increased Nrf2 translocation to the nucleus, resulting in Nrf2 target gene expression in microglia. In conclusion, DEM is a promising protective agent against NLRP3 inflammasome activation.
Insights
Diethyl Maleate (DEM) effectively reduces NLRP3 inflammasome activation in microglia by decreasing mitochondrial ROS and pyroptosis. This protective effect is mediated by DEM-induced Nrf2 activation, highlighting its therapeutic potential for neuroinflammation.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are key immune cells in the central nervous system.
- NLRP3 inflammasome activation drives neuroinflammation and neurodegenerative diseases.
- Diethyl Maleate (DEM) shows potential anti-inflammatory properties.
Purpose of the Study:
- To investigate DEM's effect on NLRP3 inflammasome activation in microglia.
- To explore DEM's impact on mitochondrial ROS and pyroptosis.
- To determine DEM's role in Nrf2 pathway activation.
Main Methods:
- Activated NLRP3 inflammasome in N9 murine microglia using LPS and ATP.
- Pretreated microglia with DEM before inflammasome activation.
- Assessed NLRP3 inflammasome activation, mitochondrial ROS levels, pyroptosis, and Nrf2 translocation.
Main Results:
- DEM pretreatment significantly attenuated NLRP3 inflammasome activation.
- DEM reduced mitochondrial ROS and pyroptotic cell death in microglia.
- DEM enhanced Nrf2 translocation to the nucleus and Nrf2 target gene expression.
Conclusions:
- DEM effectively inhibits NLRP3 inflammasome activation in microglia.
- DEM exerts protective effects by reducing oxidative stress and pyroptosis via Nrf2 activation.
- DEM is a promising therapeutic agent for neuroinflammatory conditions.
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