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Wnt1 oversees microglial activation by the Wnt/LRP5/6 receptor signaling pathway during lipopolysaccharide-mediated
Wang Qing1, Xu Hao1, Sun Xuan1
1Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Avenue, Wuhan, 430030, China.
Background:
The protective effects of autophagy-mediated microglial inflammatory regulation on diseases of the central nervous system (CNS) has been a recent field of interest. The canonical signaling pathway activated by Wnt1, the Wnt/β-catenin signaling cascade, also plays a crucial protective role in neurodegenerative diseases. However, the relationship between Wnt1/β-catenin signaling and microglial activation remains unclear. Our study focused on understanding the impact and mechanism of Wnt1 on microglial activation.
Methods And Results:
To simulate neuroinflammatory conditions in vitro, BV2 cells were exposed to 1 μg/mL lipopolysaccharide. CD86- and CD206-positive cells were identified by flow cytometry and immunofluorescence assays. Inflammatory and anti-inflammatory factors were measured using enzyme-linked immunosorbent assays. Autophagy was analyzed by expression of LC3B puncta, LC3, P62, and beclin1 expression. The inflammatory activation suppressed by rhWnt1 was restricted by DKK1, siRNA-β-catenin and siRNA-LKB1, respectively, with concomitant changes in β-catenin expression and phosphorylation of NFκB-p65, LKB1, and AMPK. Although the anti-inflammatory effect of Wnt1/LKB1 pathway was independent of β-catenin, Wnt1/LKB1 regulated β-catenin. The reduced inflammation caused by rhWnt1 is linked to its enhancement of autophagy, a process blocked by siRNA-LKB1 and 3-MA partially.
Conclusions:
The anti-inflammatory effects of Wnt1 on BV2 cells improved autophagy, a mechanism partly dependent on the β-catenin pathway or the phosphorylation of LKB1. Furthermore, the Wnt1/LKB1 pathway was activated independently of β-catenin and participated in regulating its expression. Our research unveils a previously unknown method through which Wnt1 exerts its anti-inflammatory effects, which may have a potential protective role against CNS diseases.
Insights
Wnt1 reduces neuroinflammation in microglia by enhancing autophagy, a process partly mediated by the Wnt/β-catenin pathway and LKB1 phosphorylation. This reveals a novel anti-inflammatory mechanism for Wnt1 in central nervous system diseases.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglial inflammatory regulation via autophagy shows promise for central nervous system (CNS) diseases.
- Wnt/β-catenin signaling is protective in neurodegenerative diseases, but its link to microglial activation is unclear.
- This study investigates the mechanism of Wnt1's impact on microglial activation.
Purpose of the Study:
- To elucidate the mechanism by which Wnt1 influences microglial activation.
- To explore the relationship between Wnt1/β-catenin signaling and microglial inflammatory responses.
- To understand the role of autophagy in Wnt1-mediated anti-inflammatory effects.
Main Methods:
- BV2 cells were stimulated with lipopolysaccharide to mimic neuroinflammation.
- Flow cytometry and immunofluorescence identified cell markers (CD86, CD206).
- ELISA measured inflammatory factors; autophagy was assessed via LC3B puncta and protein expression (LC3, P62, beclin1).
Main Results:
- rhWnt1 suppressed inflammatory activation, an effect blocked by DKK1, siRNA-β-catenin, and siRNA-LKB1.
- Wnt1 enhanced autophagy, which was partially inhibited by siRNA-LKB1 and 3-MA.
- The Wnt1/LKB1 pathway's anti-inflammatory effect was independent of β-catenin but regulated its expression.
Conclusions:
- Wnt1 exerts anti-inflammatory effects on microglia by enhancing autophagy.
- This mechanism involves the Wnt/β-catenin pathway and LKB1 phosphorylation.
- Wnt1 presents a novel therapeutic target for CNS diseases due to its anti-inflammatory properties.
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