Interleukin 33 regulates the autophagy of oxygen-glucose-deprivation/reoxygenation microglia via macrophage polarization
1
Central Laboratory, Affiliated Fuzhou First Hospital of Fujian Medical University, Fuzhou, Fujian, 350009, China
2
College of Life Sciences, Fujian Normal University, Fuzhou, Fujian, 350009, China
3
Department of Neurosurgery, Affiliated Fuzhou First Hospital of Fujian Medical University, Fuzhou, Fujian, 350009, China
4
Department of Respiratory Medicine, Affiliated Fuzhou First Hospital of Fujian Medical University, Fuzhou, Fujian, 350009, China
These authors had equal contribution to this work
Submission date: 2024-05-05
Final revision date: 2025-04-03
Acceptance date: 2025-04-10
Online publication date: 2026-01-23
Corresponding author
Yongkai Yang
Department of Neurosurgery, Affiliated Fuzhou First Hospital of Fujian Medical University, Fuzhou, Fujian, 350009, China
Department of Neurosurgery, Affiliated Fuzhou First Hospital of Fujian Medical University, Fuzhou, Fujian, 350009, China
KEYWORDS
ABSTRACT
Introduction:
Interleukin (IL)-33 is a pleiotropic cytokine in the immune system and inflammatory responses, which is involved in cerebral ischemia/reperfusion (I/R) injury. Evidence indicates that IL-33 plays an essential role in macrophage polarization activation, yet the potential molecular mechanisms remain elusive. This study explored the role of IL-33 in the activation of microglia/macrophage-mediated autophagy for cerebral I/R injury via in vitro experiments.
Material and Methods:
Primary macrophages were harvested from mouse bone marrow, and microglia were isolated from mouse brains. Lipopolysaccharide (LPS) and interferon (IFN-) induced polarization toward the M1 phenotype macrophage, and IL-4 induced polarization toward the M2 phenotype macrophage. Oxygen glucose deprivation/reoxygenation (OGD/R) models of primary microglia were established and co-cultured with M1 and M2 macrophages, respectively. The IL-33 knockout microglia OGD/R model and M1/M2 macrophages were co-cultured to explore the effects of IL-33 in the transformation from pro-inflammatory (M1) phenotype to anti-inflammatory (M2) phenotype against inflammation and autophagy. Microglial activation/polarization, autophagy, and inflammation were detected using western blot, immunofluorescence, and ELISA.
Results:
Compared with microglia co-cultured with M1 macrophages, microglia co-cultured with M2 macrophages showed significantly higher expression of M2 intracellular markers (STAT6, PPAR-, IRF4, CEBP-, and CD206 (p < 0.05). In contrast, the expression of STAT1 and IRF8 was significantly lower. IL-33 deficiency enhanced the anti-inflammatory phenotype of microglia/M2 macrophages. Furthermore, the expression of IL-10 and transforming growth factor (TGF-) was upregulated (p < 0.05), and the expression of IFN- was downregulated (p < 0.05) in microglia/M2 macrophages in comparison with microglia/M1 macrophages. Compared with the OGD/R model group, M1 macrophage co-culture increased Beclin-1 and ULK1 expression and reduced P62 expression, and M2 co-culture as well as IL-33 deletion reduced Beclin-1 and ULK1 expression and increased P62 expression levels.
Conclusions:
IL-33 deficiency inhibited hypoxia-induced microglia autophagy and promoted macrophage polarization toward the anti-inflammatory phenotype.
Interleukin (IL)-33 is a pleiotropic cytokine in the immune system and inflammatory responses, which is involved in cerebral ischemia/reperfusion (I/R) injury. Evidence indicates that IL-33 plays an essential role in macrophage polarization activation, yet the potential molecular mechanisms remain elusive. This study explored the role of IL-33 in the activation of microglia/macrophage-mediated autophagy for cerebral I/R injury via in vitro experiments.
Material and Methods:
Primary macrophages were harvested from mouse bone marrow, and microglia were isolated from mouse brains. Lipopolysaccharide (LPS) and interferon (IFN-) induced polarization toward the M1 phenotype macrophage, and IL-4 induced polarization toward the M2 phenotype macrophage. Oxygen glucose deprivation/reoxygenation (OGD/R) models of primary microglia were established and co-cultured with M1 and M2 macrophages, respectively. The IL-33 knockout microglia OGD/R model and M1/M2 macrophages were co-cultured to explore the effects of IL-33 in the transformation from pro-inflammatory (M1) phenotype to anti-inflammatory (M2) phenotype against inflammation and autophagy. Microglial activation/polarization, autophagy, and inflammation were detected using western blot, immunofluorescence, and ELISA.
Results:
Compared with microglia co-cultured with M1 macrophages, microglia co-cultured with M2 macrophages showed significantly higher expression of M2 intracellular markers (STAT6, PPAR-, IRF4, CEBP-, and CD206 (p < 0.05). In contrast, the expression of STAT1 and IRF8 was significantly lower. IL-33 deficiency enhanced the anti-inflammatory phenotype of microglia/M2 macrophages. Furthermore, the expression of IL-10 and transforming growth factor (TGF-) was upregulated (p < 0.05), and the expression of IFN- was downregulated (p < 0.05) in microglia/M2 macrophages in comparison with microglia/M1 macrophages. Compared with the OGD/R model group, M1 macrophage co-culture increased Beclin-1 and ULK1 expression and reduced P62 expression, and M2 co-culture as well as IL-33 deletion reduced Beclin-1 and ULK1 expression and increased P62 expression levels.
Conclusions:
IL-33 deficiency inhibited hypoxia-induced microglia autophagy and promoted macrophage polarization toward the anti-inflammatory phenotype.
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| eISSN: | 1644-4124 |
| ISSN: | 1426-3912 |
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