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Ascorbic acid 6-palmitate modulates microglia M1/M2 polarization in lipopolysaccharide-stimulated BV-2 cells via
Qian Li1,2,3, Yao Wu2,3, Xue-Shen Chen2,3
1Grade 2018, West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Sichuan, 610041, Chengdu, PR China.
Background:
Neuroinflammation-mediated microglia polarization is a major process in various central nervous system (CNS) diseases. Endoplasmic reticulum (ER) stress contributes to the inflammatory signals as well as to microglia polarization in lipopolysaccharide (LPS) induced neuroinflammation. Ascorbic acid 6-palmitate (L-AP) has been broadly used as a dietary antioxidant in foods and demonstrated a strong inhibitory effect on 5-LOX; however, the specific anti-inflammation mechanisms remain unclear. In this study, we investigated the effects and possible mechanisms of L-AP on LPS-induced neuroinflammation in BV-2 cells.
Methods:
Immortalized murine microglia cell line BV-2 cells were employed to assess the effect of L-AP to modulate microglia M1/M2 polarization in vivo, and the molecular mechanism was evaluated by qRT-PCR and Western blotting analysis. Molecular docking was used to predict the binding activity of L-AP with protein kinase R-like ER kinase (PERK).
Results:
L-AP at 62.5 µM significantly modulated LPS-induced microglia M1/M2 polarization (increases of interleukin (IL)-10 and arginase-1 (Arg-1) transcriptions) independent of cell growth. Besides, L-AP at 62.5 µM significantly down-regulated glucose-regulated protein 78 (GRP78) and CCAAT/enhancer-binding homologous protein (CHOP) mRNA levels. Similar data were shown in the tunicamycin (TM) induced ER stress cells model. Moreover, the protective effect of L-AP on TM-induced microglia M1/M2 polarization was similar to that of 4-phenyl butyric acid (4-PBA), the ER stress inhibitor. Molecular docking results indicated L-AP might directly bind with PERK, with a binding affinity of -7.7 kcal/mol. A further study unveiled that L-AP notably inhibited LPS-induced PERK/ eukaryotic initiation factor 2α (elf2α) activation.
Conclusion:
Together, this study revealed that L-AP possessed its effect on the reconstruction of microglia M1/M2 polarization balance in LPS-stimulated BV-2 cells via modulating PERK/elF2α mediated ER stress.
Insights
Ascorbic acid 6-palmitate (L-AP) rebalances microglia polarization in neuroinflammation by targeting endoplasmic reticulum (ER) stress. L-AP modulates the PERK/elF2α pathway, offering a potential therapeutic strategy for CNS diseases.
Area of Science:
- Neuroscience
- Cell Biology
- Immunology
Background:
- Microglia polarization is critical in central nervous system (CNS) diseases.
- Endoplasmic reticulum (ER) stress exacerbates neuroinflammation and microglia polarization.
- Ascorbic acid 6-palmitate (L-AP) is a dietary antioxidant with unclear anti-inflammatory mechanisms.
Purpose of the Study:
- To investigate the effects of L-AP on lipopolysaccharide (LPS)-induced neuroinflammation in BV-2 cells.
- To elucidate the underlying mechanisms of L-AP's anti-inflammatory actions, focusing on ER stress and microglia polarization.
Main Methods:
- Utilized immortalized murine microglia BV-2 cells to assess L-AP's impact on microglia M1/M2 polarization.
- Employed quantitative real-time PCR (qRT-PCR) and Western blotting to analyze molecular mechanisms.
- Performed molecular docking to predict L-AP binding with protein kinase R-like ER kinase (PERK).
Main Results:
- L-AP (62.5 µM) significantly modulated LPS-induced microglia M1/M2 polarization, increasing IL-10 and Arg-1.
- L-AP down-regulated ER stress markers (GRP78, CHOP) in both LPS and tunicamycin (TM)-induced models.
- L-AP inhibited LPS-induced PERK/eukaryotic initiation factor 2α (elF2α) activation, suggesting a direct interaction with PERK.
Conclusions:
- L-AP effectively reconstructs microglia M1/M2 polarization balance in LPS-stimulated BV-2 cells.
- The mechanism involves modulating the PERK/elF2α-mediated ER stress pathway.
- L-AP demonstrates potential as a therapeutic agent for neuroinflammatory conditions.
