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Updated: May 15, 2026

Ferric Chloride-induced Murine Thrombosis Models
Published on: September 5, 2016
Theaflavin protects against ischemic stroke by inhibiting PLA2G4C-driven lipid remodeling and ferroptosis
Yang Yang1, Quandan Tan2, Meiling Yao3
1Department of Neurology, The Affiliated Hospital of Southwest Medical University, Luzhou, China.
Background:
Ferroptosis contributes to secondary brain injury after ischemic stroke, but upstream phospholipid remodeling mechanisms remain unclear.
Purpose:
We investigated whether PLA2G4C promotes post-ischemic ferroptosis and whether theaflavin confers neuroprotection by targeting this pathway.
Study Design:
Combined in vivo and in vitro mechanistic study using rat MCAO/R and OGD/R models.
Methods:
RNA sequencing and siRNA-mediated knockdown assessed PLA2G4C function in MCAO/R rats and OGD/R neurons. Virtual screening identified theaflavin as a candidate modulator; surface plasmon resonance (SPR) compared binding affinities of three shortlisted compounds to recombinant PLA2G4C; cellular thermal shift assay (CETSA) assessed intracellular target engagement; LC-MS/MS lipidomics characterized PE-AA remodeling.
Results:
PLA2G4C was significantly upregulated in ischemic brain tissue, predominantly in neurons, and was associated with ferroptosis-related transcriptional signatures. Pla2g4c silencing reduced infarct size and improved neurological outcomes, effects reversed by RSL3. PLA2G4C knockdown reduced PE-AA accumulation, attenuated ACSL4 upregulation, and preserved GPX4. SPR confirmed theaflavin as the strongest binder among three candidates (KD = 243 nM vs. 2.11 μM and 8.69 μM for the comparators). CETSA supported intracellular target engagement. Theaflavin reduced PLA2G4C abundance and PE-AA content, attenuated lipid peroxidation, and improved neuronal survival; PE-AA supplementation or PLA2G4C overexpression diminished these benefits.
Conclusion:
PLA2G4C drives post-ischemic ferroptosis through PE-AA accumulation. SPR and CETSA support direct binding and intracellular engagement of PLA2G4C by theaflavin, providing mechanistic evidence for its anti-ferroptotic and neuroprotective effects.
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