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Published on: January 30, 2014
Punicalagin Attenuates LPS-Induced Inflammation and ROS Production in Microglia by Inhibiting the MAPK/NF-κB
Jung Lo1,2, Ching-Chih Liu3,4, Yueh-Shan Li3
1Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan.
Purpose:
Neurodegenerative diseases are associated with neuroinflammation along with activation of microglia and oxidative stress, but currently lack effective treatments. Punicalagin is a natural bio-sourced product that exhibits anti-inflammatory effects on several chronic diseases; however, the anti-inflammatory and anti-oxidative effects on microglia have not been well examined. This study aimed to investigate the effects of punicalagin on LPS-induced inflammatory responses, NLRP3 inflammasome activation, and the production of ROS using murine microglia BV2 cells.
Methods:
BV2 cells were pre-treated with punicalagin following LPS treatment to induce inflammation. The secretion of NO and PGE2 was analyzed by Griess reagent and ELISA respectively, while the expressions of iNOS, COX-2, STAT3, ERK, JNK, and p38 were analyzed using Western blotting, the production of IL-6 was measured by ELISA, and the activity of NF-κB was detected using promoter reporter assay. To examine whether punicalagin affects NLRP3 inflammasome activation, BV2 cells were stimulated with LPS and then treated with ATP or nigericin. The secretion of IL-1β was measured by ELISA. The expressions of NLRP3 inflammasome-related proteins and phospho IκBα/IκBα were analyzed using Western blotting. The production of intracellular and mitochondrial ROS was analyzed by flow cytometry.
Results:
Our results showed that punicalagin attenuated inflammation with reduction of pro-inflammatory mediators and cytokines including iNOS, COX-2, IL-1β, and reduction of IL-6 led to inhibition of STAT3 phosphorylation by LPS-induced BV2 cells. Punicalagin also suppressed the ERK, JNK, and p38 phosphorylation, attenuated NF-κB activity, inhibited the activation of the NLRP3 inflammasome, and reduced the production of intracellular and mitochondrial ROS by LPS-induced BV2 cells.
Conclusion:
Our results demonstrated that punicalagin attenuated LPS-induced inflammation through suppressing the expression of iNOS and COX-2, inhibited the activation of MAPK/NF-κB signaling pathway and NLRP3 inflammasome, and reduced the production of ROS in microglia, suggesting that punicalagin might have the potential in treating neurodegenerative diseases.
Insights
Punicalagin reduces neuroinflammation and oxidative stress in microglia by inhibiting key inflammatory pathways. This natural compound shows potential for treating neurodegenerative diseases by calming activated microglia.
Area of Science:
- Neuroscience
- Immunology
- Pharmacology
Background:
- Neurodegenerative diseases involve neuroinflammation, microglial activation, and oxidative stress.
- Current treatments for these conditions are limited.
- Punicalagin, a natural product, has shown anti-inflammatory effects in other diseases.
Purpose of the Study:
- To investigate punicalagin's effects on lipopolysaccharide (LPS)-induced inflammatory responses in murine microglia BV2 cells.
- To examine punicalagin's impact on NLRP3 inflammasome activation and reactive oxygen species (ROS) production.
- To evaluate punicalagin's potential therapeutic role in neuroinflammation.
Main Methods:
- BV2 cells were treated with punicalagin and LPS to induce inflammation.
- Inflammatory mediators (NO, PGE2, IL-6, IL-1β), protein expressions (iNOS, COX-2, STAT3, ERK, JNK, p38), and signaling pathway activity (NF-κB, NLRP3 inflammasome) were analyzed.
- Intracellular and mitochondrial ROS production was measured via flow cytometry.
Main Results:
- Punicalagin significantly reduced pro-inflammatory mediators and cytokines (iNOS, COX-2, IL-1β, IL-6).
- It inhibited STAT3, ERK, JNK, and p38 phosphorylation, and attenuated NF-κB activity.
- Punicalagin suppressed NLRP3 inflammasome activation and decreased intracellular and mitochondrial ROS production.
Conclusions:
- Punicalagin effectively attenuates LPS-induced inflammation and oxidative stress in microglia.
- It inhibits key inflammatory signaling pathways, including MAPK/NF-κB and NLRP3 inflammasome.
- Punicalagin demonstrates potential as a therapeutic agent for neurodegenerative diseases.
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