Related Experiment Video
Updated: Apr 18, 2026
Three-dimensional Confocal Analysis of Microglia/macrophage Markers of Polarization in Experimental Brain Injury
Published on: September 4, 2013
IFITM1 promotes microglial polarization and NLRP3 inflammasome activation in ischemic stroke through up-regulating
1Department of Neurology, Xi'an International Medical Center Hospital, Xi'an, 710010 Shaanxi China.
Abstract:
Microglial polarization and the NLRP3 inflammasome play critical roles in the pathology of ischemic stroke (IS). Regulation of microglial polarization towards M2 type has become a potential therapy approach for M2 type has become a potential therapy approach for treating IS. The current study demonstrated IFITM1 was markedly upregulated in microglia in the Distal middle cerebral artery (dMCAO) model. This research sought to assess the protective potential of IFITM1 knockdown in IS and to study its downstream regulatory mechanisms. The dMCAO mice model and oxygen-glucose deprivation/reoxygenation (OGD/R) BV2 cell model were constructed to imitate IS injury. The GSE148350 dataset was utilized to screen for differentially expressed genes in IS. Immunofluorescence, western blotting, and ELISA were utilized to evaluated activation level of NLRP3 inflammasome. RT-qPCR was applied to evaluated gene expression associated with the M1/M2 phenotypes and glycolysis of microglia. After treatment with dMCAO and OGD/R, IFITM1 expression was significantly upregulated. Knockdown of IFITM1 significantly inhibited M1 markers and promoted M2 markers of microglia. Additionally, knockdown of IFITM1 obviously inhibited the activation of the NLRP3 inflammasome and glycolysis. The expression of c-FOS was markedly raised following OGD/R, but was obviously inhibited by knockdown of IFITM1. Rescue experiments showed that knockdown of IFITM1 inhibited the NLRP3 inflammasome activation and M1 polarization of microglia by suppressing c-FOS expression. Knockdown of IFITM1 alleviated brain infarction, brain edema, and neurological damage in dMCAO mice. These results suggest that the absence of IFITM1 alleviates brain injury induced by dMCAO, inhibiting NLRP3 inflammasome activation and glycolysis of microglia through suppressing c-FOS expression.
Supplementary Information:
The online version contains supplementary material available at 10.1007/s10616-025-00832-5.
Insights
IFITM1 knockdown protects against ischemic stroke by reducing microglial M1 polarization and NLRP3 inflammasome activation. This intervention alleviates brain injury via suppressing c-FOS expression and glycolysis.
Area of Science:
- Neuroscience
- Immunology
- Molecular Biology
Background:
- Ischemic stroke (IS) pathology involves microglial polarization and NLRP3 inflammasome activation.
- Shifting microglia to the M2 phenotype is a potential therapeutic strategy for IS.
- IFITM1 is upregulated in microglia during IS, suggesting a role in the disease process.
Purpose of the Study:
- To investigate the protective effects of IFITM1 knockdown in IS.
- To elucidate the downstream mechanisms regulated by IFITM1 in IS.
Main Methods:
- Established distal middle cerebral artery occlusion (dMCAO) mice and oxygen-glucose deprivation/reoxygenation (OGD/R) BV2 cell models.
- Utilized the GSE148350 dataset to identify differentially expressed genes in IS.
- Employed immunofluorescence, western blotting, ELISA, and RT-qPCR to assess microglial activation, NLRP3 inflammasome, gene expression, and glycolysis.
Main Results:
- IFITM1 expression was significantly upregulated in dMCAO and OGD/R models.
- IFITM1 knockdown suppressed M1 microglial markers, promoted M2 markers, and inhibited NLRP3 inflammasome activation and glycolysis.
- Knockdown of IFITM1 reduced brain infarction, edema, and neurological deficits in dMCAO mice, mediated by c-FOS suppression.
Conclusions:
- IFITM1 absence alleviates dMCAO-induced brain injury.
- IFITM1 inhibition of NLRP3 inflammasome activation and microglial glycolysis occurs through c-FOS suppression.
- Targeting IFITM1 presents a potential therapeutic avenue for ischemic stroke.
