Metabolomics Analysis Revealed That SXZ Ameliorates Cognitive Impairment in Naturally Aged Rats by Promoting Glutathione Metabolism and Inhibiting the Ferroptosis Pathway

ABSTRACT Objective This study aimed to examine the potential neuroprotective effects of SXZ on cognitive impairment in naturally aging rats and to explore the underlying molecular mechanisms. Methods Eighteen‐month‐old Sprague–Dawley rats were used as a natural aging model and administered Shenxian Zhou (SXZ) for 6 months. Cognitive function was assessed using behavioral evaluations. Hematoxylin–eosin, Nissl, and immunofluorescence staining for NeuN were performed to evaluate neuronal pathology in aging rats. Metabolomics analysis was performed to characterize hippocampal metabolites, followed by molecular validation. Results SXZ attenuated age‐related cognitive decline and reduced hippocampal pathological and neuronal damage in naturally aging rats. SXZ‐H significantly modulated glutathione metabolism and ferroptosis‐associated signaling pathways. Transmission electron microscopy revealed mitochondrial morphological changes consistent with ferroptosis in hippocampal neurons of naturally aging rats. Molecular biology experiments further indicated that, subsequent to SXZ‐H intervention, lipid peroxidation and oxidative stress levels were significantly reduced. Furthermore, expression levels of p53 and Keap1 genes and proteins were significantly downregulated, whereas the expression of SLC7A11, GPX4, Nrf2, and HO‐1 genes and proteins was notably upregulated. Conclusion SXZ‐H effectively mitigates cognitive impairment in naturally aging rats, an effect that may be mediated by promoting glutathione metabolism and regulating key ferroptosis‐related proteins, thereby inhibiting neuronal ferroptosis.