KLF4 promotes sevoflurane-induced neurotoxicity by suppressing FOXO1 expression in neonatal rats

Background: Exposure to anesthetics such as sevoflurane (SEV) during early development has been associated with neurotoxicity. Krüppel-like factor 4 (KLF4), a transcription factor implicated in neuronal injury, has been suggested to play a role in this process. However, the underlying mechanisms remain inadequately characterized. Methods: In vivo, neonatal rats were intravenously administered adeno-associated virus (AAV) constructs with or without 3% SEV exposure. Their neurological functions were evaluated, and hippocampal tissues were harvested for molecular and histopathological analyses, including NISSL staining, hematoxylin and eosin (H&E) staining, modified Bielschowsky silver staining, Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR), western blotting, Terminal deoxynucleotidyl transferase dUTP Nick End Labeling (TUNEL) assay, oxidative stress assessment and dihydroethidium (DHE) staining. Behavioral assessment was conducted using the Morris water maze (MWM). In vitro, HT22 hippocampal neurons were transfected with KLF4-targeting Small Interfering RNAs (siRNAs) and exposed to 3% SEV to assess changes in cell morphology, apoptosis and oxidative stress. Results: Sevoflurane exposure led to significant neurotoxicity in neonatal rats, characterized by cognitive impairment, neuronal apoptosis, and oxidative stress, accompanied by increased KLF4 and decreased Forkhead Box Protein O1 (FOXO1) expression. The knockdown of KLF4 using Short Hairpin RNA (shRNA) reversed these effects, improving cognitive performance and reducing neuronal damage and oxidative stress. In vitro, KLF4 silencing was found to mitigate SEV-induced cellular injury in HT22 cells. Notably, KLF4 knockdown upregulated FOXO1 expression, which further attenuated SEV-induced apoptosis and oxidative stress. Conclusions: These findings indicate that KLF4 aggravates sevoflurane-induced neurotoxicity in neonatal rats by downregulating FOXO1.

Neonatal rats; KLF4; FOXO1; Sevoflurane; Neurotoxicity

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