Acupuncture Inhibits Ferroptosis to Attenuate Cerebral Ischemia-Reperfusion Injury Through FTO-Mediated m6A Modification of ACSL4.

Abstract

Acupuncture, a traditional Chinese medical intervention, is extensively utilized in the management of ischemic stroke across its various stages. However, the precise mechanisms underlying its therapeutic action are not fully elucidated. The present study sought to investigate the therapeutic effects and unravel the potential mechanisms of Xingnao Kaiqiao (XNKQ) acupuncture in ameliorating cerebral ischemia-reperfusion injury (CIRI). We established a rat model of middle cerebral artery occlusion (MCAO) and administered both Xingnao Kaiqiao (XNKQ) acupuncture and non-acupoint acupuncture as interventions. Neurobehavioral scoring and TTC staining were employed to evaluate the efficacy of acupuncture in improving CIRI. Our findings indicated that XNKQ acupuncture significantly enhanced neurological function and decreased the volume of cerebral infarction in rats. Western blot, quantitative real-time PCR (RT-qPCR), dot blot, and kit assays were utilized to assess the impact of acupuncture on ferroptosis and m6A methylation. The data revealed that XNKQ acupuncture diminished the levels of ACSL4 and the ferroptosis-associated markers MDA and Fe, concurrently reducing global m6A levels and selectively upregulating FTO expression. Subsequently, FTO overexpression and knockdown vectors were administered to MCAO rats, and the interaction between FTO and ACSL4 was confirmed through bioinformatics analysis and methylated RNA immunoprecipitation (MeRIP). The outcomes demonstrated that FTO could modulate the expression of ACSL4 by regulating the m6A methylation of ACSL4 mRNA. Further acupuncture interventions revealed that acupuncture suppresses ACSL4 expression by increasing FTO expression, thereby reducing m6A methylation of ACSL4, downregulating MDA and Felevels, and inhibiting ferroptosis. Collectively, these results suggest that acupuncture mitigates CIRI by modulating the upregulation of FTO, leading to reduced m6A levels of ACSL4 and the inhibition of ferroptosis.