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Comprehensive Evaluation of YJ-2 as a PAD4 Inhibitor in Alleviating Ischemic Brain Injury: From NETs-Induced
Yijiang Jia1,2, Ayijiang Taledaohan3, Kai Wang1,2
1Department of Medicinal Chemistry, School of Pharmaceutical Sciences of Capital Medical University, Beijing, China.
Aims:
To evaluate the neuroprotective potential of YJ-2, a novel peptidylarginine deiminase 4 (PAD4) inhibitor, against ischemia/reperfusion brain injury by targeting neutrophil extracellular trap (NET) formation.
Methods:
In vitro, a NETs-induced injury model was established using SH-SY5Y and bEnd.3 cells. YJ-2's effects on viability, apoptosis, oxidative stress, and barrier permeability were assessed via CCK-8, flow cytometry, and FITC-dextran assays. In vivo, a rat middle cerebral artery occlusion/reperfusion (MCAO/R) model received YJ-2 (10 μmol/kg) intravenously. Outcomes included infarct volume (TTC staining), neurological score, neuronal apoptosis (TUNEL), and oxidative markers (ELISA). PAD4 activity and histone H3 citrullination (H3cit) were examined by western blot and immunofluorescence.
Results:
YJ-2 reduced NET-mediated neuronal death and oxidative stress in vitro, and improved endothelial barrier integrity. In MCAO/R rats, YJ-2 significantly lowered infarct volume (44.2% → 30.6%), improved neurological function, and suppressed apoptosis. It also decreased PAD4 and H3cit expression in ischemic brain tissue, confirming target engagement.
Conclusion:
YJ-2, by preserving blood-brain barrier (BBB) integrity and reducing neuronal apoptosis, highlights its therapeutic potential for ischemic stroke.
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