Related Experiment Video
Updated: Jan 10, 2026
Herbal Munziq Ameliorates Myocardial Ischemia-Reperfusion Injury by Inhibiting Inflammation
Published on: January 10, 2025
TNFAIP3 alleviates cerebral ischemia-reperfusion injury by inhibiting M1 microglia polarization via deubiquitination
Wenya Bai1, Shixuan Liu2, Guilin Zhou1
1Department of Anesthesiology, The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, P. R. China.
Background:
Microglia polarization plays a crucial role in the progression of cerebral ischemia-reperfusion injury (CIRI), but the mechanisms remain largely undefined. The preset study aimed to investigate the mechanism of microglia polarization following CIRI.
Methods:
CIRI was modeled in C57BL/6J mice through middle cerebral artery occlusion-reperfusion and in BV2 cells via oxygen and glucose deprivation/reoxygenation. Reverse transcription-quantitative PCR, western blotting, flow cytometry and fluorescence staining were used to detect the expression levels of key proteins associated with microglia polarization, as well as the expression of TNFAIP3 and RACK1. The interaction between TNFAIP3 and RACK1 was verified by co-immunoprecipitation. TNFAIP3 or RACK1 gene interference (overexpression and/or silencing) was employed to examine the role of the TNFAIP3/RACK1 axis in microglia polarization following CIRI.
Results:
The results revealed that Arg-1 expression decreased, inducible nitric oxide synthase expression increased and TNFAIP3 was upregulated 24 h after CIRI. Furthermore, TNFAIP3 interacted with RACK1 to deubiquitinate and increase the expression of RACK1. These results indicate that knocking down either TNFAIP3 or RACK1 promotes microglia M1 polarization, and overexpression of RACK1 can promote microglia M2 polarization. RACK1 exerts its neuroprotective effects through NF-κB, as demonstrated by the use of NF-κB inhibitors.
Conclusion:
The present findings indicate that TNFAIP3 inhibits M1 microglial polarization via deubiquitination of RACK1 after CIRI, RACK1 exerts its effects through NF-κB.
Insights
TNFAIP3 inhibits M1 microglia polarization by deubiquitinating RACK1 after cerebral ischemia-reperfusion injury. RACK1 then exerts neuroprotective effects via NF-κB signaling, promoting M2 polarization.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia polarization is critical in cerebral ischemia-reperfusion injury (CIRI) progression.
- The precise mechanisms regulating microglia polarization in CIRI are not fully understood.
Purpose of the Study:
- To elucidate the mechanism of microglia polarization in the context of CIRI.
- To investigate the role of the TNFAIP3/RACK1 axis in regulating microglial responses.
Main Methods:
- Cerebral ischemia-reperfusion injury (CIRI) was induced in mice and BV2 cells.
- Quantitative PCR, Western blotting, and flow cytometry assessed protein expression and microglia polarization markers.
- Co-immunoprecipitation and gene interference (overexpression/silencing) explored the TNFAIP3-RACK1 interaction and its functional impact.
Main Results:
- TNFAIP3 expression increased post-CIRI, interacting with RACK1 to increase RACK1 expression via deubiquitination.
- Silencing TNFAIP3 or RACK1 promoted M1 microglia polarization; RACK1 overexpression promoted M2 polarization.
- RACK1 mediated its neuroprotective effects through the NF-κB pathway.
Conclusions:
- TNFAIP3 inhibits M1 microglia polarization by deubiquitinating RACK1 in CIRI.
- RACK1 plays a key role in neuroprotection following CIRI, acting through the NF-κB pathway.
