Related Experiment Video
Updated: Jul 11, 2025
Assessing Retinal Microglial Phagocytic Function In Vivo Using a Flow Cytometry-based Assay
Published on: October 18, 2016
Docosahexaenoic Acid (DHA) Reduces LPS-Induced Inflammatory Response Via ATF3 Transcription Factor and Stimulates
Katarzyna Wieczorek-Szukala1, Monika Markiewicz2, Anna Walczewska1
1Department of Cell-to-Cell Communication, Medical University of Lodz, Lodz, Poland.
Background/Aims:
Microglial cells play a crucial role in the development of neuroinflammation in response to harmful stimuli, such as infection, ischemia or injury. Their chronic activation, however, is associated with a progression of neurodegenerative diseases. Therefore, looking for potential factors limiting microglial activation, the effect of docosahexaenoic acid (DHA) on the inflammatory response and TREM2-dependent phagocytic activity in microglia was investigated.
Methods:
In LPS-induced primary microglia preincubated with DHA, or without preincubation the expression of ATF3 and TREM2 genes and TREM2, Syk, Akt proteins were determined by RT-PCR and WB, respectively. Cell viability was assayed by MTT and cytokine and chemokine expression was determined by the Proteome Profiler assay. Moreover, the phagocytic activity of microglia was assayed using immunofluorescence.
Results:
We found that DHA significantly increased the expression of ATF3 , and decreased the levels of CINC-1, CINC-2αβ, CINC-3 chemokines, IL-1α and IL-1β cytokines, and ICAM-1 adhesion protein. Additionally, preincubation of microglia with DHA resulted in increased Src/Syk kinases activation associated with increased phagocytic microglia activity.
Conclusion:
These findings indicate that DHA efficiently inhibits ATF3-dependent release of proinflammatory mediators and enhances phagocytic activity of microglia. The study provides a new mechanism of DHA action in reactive microglia, which may help limit neuronal damage caused by the pro-inflammatory milieu in the brain.
Insights
Docosahexaenoic acid (DHA) reduces neuroinflammation by inhibiting pro-inflammatory mediators and enhancing microglial phagocytic activity. This suggests DHA may protect the brain from damage in neurodegenerative diseases.
Area of Science:
- Neuroscience
- Immunology
- Biochemistry
Background:
- Microglial cells are key in neuroinflammation, but chronic activation drives neurodegeneration.
- Identifying factors to limit microglial activation is crucial for treating neurodegenerative diseases.
Purpose of the Study:
- To investigate the effects of docosahexaenoic acid (DHA) on microglial inflammatory response.
- To examine DHA's impact on TREM2-dependent phagocytic activity in microglia.
Main Methods:
- Primary microglia were treated with DHA and lipopolysaccharide (LPS).
- Gene and protein expression (ATF3, TREM2, TREM2, Syk, Akt) were analyzed via RT-PCR and Western Blot.
- Cytokine/chemokine profiles and phagocytic activity were assessed.
Main Results:
- DHA increased ATF3 expression and enhanced microglial phagocytosis.
- DHA significantly reduced pro-inflammatory mediators (chemokines, cytokines, ICAM-1).
- DHA promoted Src/Syk kinase activation.
Conclusions:
- DHA effectively inhibits the release of pro-inflammatory mediators via ATF3.
- DHA enhances microglial phagocytic capacity.
- DHA offers a potential therapeutic mechanism to mitigate neuroinflammation and neuronal damage.
More Related Videos
11:48Isolation Protocol of Mouse Monocyte-derived Dendritic Cells and Their Subsequent In Vitro Activation with Tumor Immune Complexes
Published on: May 31, 2018
09:22High Throughput Fluorometric Technique for Assessment of Macrophage Phagocytosis and Actin Polymerization
Published on: November 27, 2014
Related Concept Videos
01:38Inflammation
01:17Phagocytosis of Apoptotic Cells
Normal cells contain receptors that prevent them from being recognized...
01:20The JAK-STAT Signaling Pathway