Related Experiment Video
Updated: Aug 7, 2026
LPS and ATP-induced Death of PMA-differentiated THP-1 Macrophages and its Validation
Published on: May 3, 2024
Lactylation-driven YTHDF3 promotes microglial pyroptosis in sepsis-associated encephalopathy via the Usp9x/NLRP3 Axis
Fengzhen Huang1, Jiping Yi2, Tieqiao Zhou3
1Department of Neurology, The First People's Hospital of Chenzhou, The First School of Clinical Medicine, Xiangnan University, Chenzhou, 423000, Hunan, China; Department of Functional Neurology, & Medical Research Center, The Affiliated Guangdong Second Provincial General Hospital of Jinan University, 510317, Guangzhou, China; Department of Neurology, The First People's Hospital of Chenzhou, Hengyang Medical School, University of South China, Chenzhou, 423000, Hunan, China.
Background:
YTHDF3 expression is up-regulated in sepsis-associated encephalopathy (SAE), yet its precise function remains to be fully elucidated. This study aims to investigate the role of YTHDF3 in microglial pyroptosis during SAE and to explore its upstream and downstream regulatory mechanisms.
Methods:
Lipopolysaccharide (LPS) induced SAE models were established in mice and primary microglia for the assessment of YTHDF3 and pyroptosis levels. Downstream targets of YTHDF3 were identified through RNA sequencing and RNA immunoprecipitation. Ultimately, chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR) was utilized to confirm H3K18la enrichment at the YTHDF3 promoter.
Results:
In both in vitro and in vivo models of SAE, YTHDF3 expression was significantly upregulated. Crucially, YTHDF3 knockdown effectively attenuated NLRP3-mediated microglial pyroptosis, thereby alleviating neuroinflammation and restoring cognitive function in SAE mice. Mechanistically, Usp9x acted as a direct downstream target of YTHDF3 to mediate the deubiquitination of NLRP3, which in turn augmented microglial pyroptosis in SAE. Furthermore, the transcriptional upregulation of YTHDF3 was promoted by H3K18la enrichment at its promoter. Inhibition of the lactylation-driven YTHDF3/Usp9x/NLRP3 signaling axis attenuates microglial pyroptosis and ameliorates cognitive dysfunction in SAE models.
Conclusion:
Lactylation-driven YTHDF3 promotes microglial pyroptosis and cognitive impairment in SAE by targeting the Usp9x/NLRP3 axis. Our findings highlight this axis as a potential therapeutic target for SAE.
Insights
YTHDF3 promotes microglial pyroptosis and cognitive decline in sepsis-associated encephalopathy (SAE) by activating the Usp9x/NLRP3 pathway. Targeting this axis offers a potential therapeutic strategy for SAE.
Area of Science:
- Neuroscience
- Immunology
- Molecular Biology
Background:
- Sepsis-associated encephalopathy (SAE) involves increased YTHDF3 expression, but its role in microglial pyroptosis is unclear.
- Investigating YTHDF3's function in SAE microglial pyroptosis and its regulatory mechanisms is crucial.
Purpose of the Study:
- To elucidate the role of YTHDF3 in microglial pyroptosis during SAE.
- To identify upstream and downstream regulatory mechanisms of YTHDF3 in SAE.
Main Methods:
- Established lipopolysaccharide (LPS)-induced SAE models in mice and primary microglia.
- Assessed YTHDF3 and pyroptosis levels.
- Identified YTHDF3 targets via RNA sequencing and RNA immunoprecipitation.
- Confirmed H3K18la enrichment at the YTHDF3 promoter using ChIP-qPCR.
Main Results:
- YTHDF3 expression was significantly upregulated in both in vitro and in vivo SAE models.
- YTHDF3 knockdown attenuated NLRP3-mediated microglial pyroptosis, reducing neuroinflammation and improving cognitive function in SAE mice.
- YTHDF3 targeted Usp9x, promoting NLRP3 deubiquitination and augmenting pyroptosis; H3K18la enrichment upregulated YTHDF3 transcription.
- Inhibition of the lactylation-driven YTHDF3/Usp9x/NLRP3 axis ameliorated SAE-related cognitive dysfunction.
Conclusions:
- Lactylation-driven YTHDF3 exacerbates microglial pyroptosis and cognitive impairment in SAE via the Usp9x/NLRP3 axis.
- The YTHDF3/Usp9x/NLRP3 axis represents a promising therapeutic target for SAE.