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Updated: Sep 27, 2026
Revealing the Ferroptotic Phenotype of Medulloblastoma
Published on: March 15, 2024
C21 ameliorates microglial ferroptosis and neuronal apoptosis after ischemic brain injury by activating the
Yijiu Wu1, Xinqian Zhou1, Xufang Huo2
1School of Anesthesiology, Guizhou Medical University, Guiyang 550004, Guizhou Province, China.
Background:
Microglial ferroptosis is a key risk factor promoting neuroinflammation after ischemic brain injury (IBI). Although Compound 21 (C21) has neuroprotective and anti-inflammatory properties, its role in modulating microglial ferroptosis in IBI remains unexplored.
Methods:
IBI was induced in C57BL/6 mice via transient middle cerebral artery occlusion (tMCAO). Parallelly, an in vitro oxygen-glucose deprivation (OGD) model was established using BV2 microglial cells to mimic ischemic conditions. Gene and protein expression levels were quantified using quantitative real-time polymerase chain reaction (qRT-PCR), western blot, and immunofluorescence staining. Cell viability and apoptosis were assessed using cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. The interactions between methyltransferase-like 3 (METTL3), ubiquitin-specific peptidase 18 (USP18), and fatty acid synthase (FASN) were analyzed by RNA immunoprecipitation (RIP) and co-immunoprecipitation (Co-IP) assays. The ubiquitination of FASN was assessed by immunoprecipitation (IP) assay. The N6-methyladenosine (m6A) modification of USP18 mRNA was analyzed by m6A RNA immunoprecipitation (Me-RIP) and dual-luciferase reporter assays.
Results:
C21 treatment alleviated neuroinflammation and neuronal apoptosis in IBI models by inhibiting microglial ferroptosis. Mechanistically, C21 upregulated USP18 via m6A-dependent mRNA stabilization through increasing METTL3 expression. In addition, C21 stabilized FASN protein through promoting USP18-mediated deubiquitination. Functional experiments further demonstrated that C21 ameliorated microglial ferroptosis and neuronal apoptosis after IBI by activating the METTL3/USP18/FASN axis.
Conclusion:
C21 ameliorated neuroinflammation and neuronal apoptosis following IBI by inhibiting microglial ferroptosis through activating the METTL3/USP18/FASN axis. Our findings provide mechanistic insights into the protective role of C21 in IBI and establish a preclinical foundation for further evaluation of its therapeutic potential.