Objective To investigate the effects of hypoxia-preconditioned human dental pulp stem cells (H-hDPSCs) transplantation on neuronal ferroptosis in neonatal rats with hypoxic-ischemic brain damage (HIBD), in order to provide novel therapeutic insights for neonatal HIBD. Methods A total of 64 7-day-old Sprague Dawley rats were randomly assigned to four groups: sham-operated (Sham), HIBD, normoxic-cultured hDPSCs (N-hDPSCs), and hypoxia-preconditioned hDPSCs (H-hDPSCs), with 16 rats in each group.The HIBD model was established using the Rice-Vannucci method.N-hDPSCs or H-hDPSCs were transplanted into the lateral ventricle 24 hours post-modeling.At 72 hours post-transplantation, histopathological changes were assessed by hematoxylin-eosin (HE) and Nissl staining, serum and cortical iron levels were measured by ELISA.Expression levels of ACSL4, GPX4, and SLC7A11 mRNA in the cerebral cortex detected by real-time quantitative fluorescence polymerase chain reaction (qRT-PCR). Results The results of HE and Nissl staining showed that compared with the Sham group, HIBD group showed neuronal disarray, pyknotic nuclei, and reduced Nissl bodies, while both hDPSCs groups exhibited attenuated damage.Compared with the Sham group, serum iron and iron content of the cortical tissue on the injured side were as the followings in the remaining three groups: HIBD>N-hDPSCs>Sham (F=79.565, 121.065, P<0.001).In terms of ferroptosis markers: GPX4+/SLC7A11+ neurons were as the followings: Sham>H-hDPSCs>HIBD (F=30.896, 75.089, P<0.001), ACSL4+ neurons were as the followings: HIBD>N-hDPSCs>Sham (F=87.724, P<0.001).H-hDPSCs showed higher GPX4 or SLC7A11 and lower ACSL4 expression than N-hDPSCs at both mRNA (qRT-PCR) and protein (Western blot) levels (P<0.001). Conclusions Transplantation of hDPSCs mitigates HIBD by suppressing neuronal ferroptosis, with hypoxia-preconditioned cells demonstrating superior therapeutic effects through regulation of ferroptosis-related pathways (GPX4/SLC7A11 upregulation and ACSL4 downregulation).
Key words
hypoxia /
human dental pulp stem cells /
neonatal rats /
hypoxic-ischemic brain damage /
ferroptosis
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