White matter injury in neonatal rats is attenuated by GsMTx4 inhibiting oligodendrocyte precursor cell ferroptosis via the PIEZO1/GCLC signaling pathway.

BACKGROUND AND

objectivesThe mechanosensitive ion channel PIEZO1 has been recognized as a therapeutic target for a range of neurological disorders. Nevertheless, its involvement and underlying mechanisms in neonatal white matter injury (WMI) remain inadequately understood. This investigation was conducted to explore the role of PIEZO1 and its associated mechanisms in WMI using both rat and cellular models.

methodsA rat model of WMI was developed through the lateral ventricular administration of lipopolysaccharide (LPS), while an in vitro WMI model was developed by preconditioning oligodendrocyte precursor cells (OPCs) with LPS. Following the administration of the PIEZO1 inhibitor GsMTx4 in both in vivo and in vitro WMI models, histopathological alterations in brain tissue were evaluated via hematoxylin and eosin staining. Western blotting was utilized to evaluate the protein levels of PIEZO1, inflammatory cytokines (IL-18 and TNF-α), and ferroptosis-associated markers (ACSL4, NOX1, SLC7A11, and GPX4). The expression of myelin basic protein and PIEZO1 was further examined through immunofluorescence analysis. Moreover, ultrastructural modifications in OPCs mitochondria were investigated using transmission electron microscopy. RNA sequencing to detect differences in ferroptosis gene expression in OPCs of different treatments; The Cell Counting Kit-8 (CCK-8) measures the optical density (OD) of OPCs to determine the IC50 of LPS, and the ROS kit measures the ROS level in OPCs; Wound healing assays for OPCs multiplication and mobility; The open-field experiment and the Morris water maze experiment evaluated the anxiety-like behavior, learning, and memory abilities of rats in each group.

resultsThe administration of GsMTx4 was observed to mitigate pathological damage and inflammatory responses in WMI, alongside promoting OPCs proliferation. Additionally, OPCs ferroptosis was inhibited by GsMTx4, potentially due to the upregulation of the glutamate-cysteine ligase catalytic subunit.

conclusionsThis study highlights that GsMTx4 alleviates WMI pathological damage by suppressing OPCs ferroptosis, a process possibly mediated via the PIEZO1/GCLC signaling pathway. IMPACT: GsMTx4 protects against neonatal white matter injury (WMI) by inhibiting oligodendrocyte precursor cell (OPC) ferroptosis and inflammation, mediated through the PIEZO1/GCLC signaling pathway. This is the first study demonstrating that GsMTx4 alleviates WMI by targeting the PIEZO1/GCLC pathway to suppress OPC ferroptosis, revealing a novel mechanistic link in WMI pathogenesis. This work identifies PIEZO1 inhibition as a promising therapeutic strategy for neonatal WMI and provides crucial mechanistic insights for developing targeted neuroprotective interventions.