Ferroptosis is an iron-dependent form of oxidative cell death involved in the pathogenesis of ischemic stroke. Electroacupuncture (EA) is an effective therapeutic intervention for ischemic stroke with antioxidant effects. However, the potential connection between EA and ferroptosis, as well as the underlying mechanisms, are largely unexplored. Here, we explored the vital role of EA in ferroptosis-mediated ischemic insult and the underlying mechanisms in the male rats with middle cerebral artery occlusion/reperfusion (MCAO/R). To determine whether preserved TFEB activity is responsible for EA-mediated inhibition of ferroptosis and lysosomal dysfunction in CIR, we used shRNA-mediated knockdown of TFEB in neurons. The results showed that EA scavenged lipid peroxidation, alleviated neuronal ferroptosis, and ferritinophagy in cerebral ischemia-reperfusion (CIR). EA also suppressed lysosomal dysfunction and preserved transcription factor EB (TFEB) activation, which was at least partially mediated through mTORC1 (mechanistic target of rapamycin kinase complex 1). The loss of TFEB counteracted EA-mediated inhibition of ferroptosis and lysosomal dysfunction in CIR. Furthermore, EA significantly alleviated CIR-induced neuronal injury in a TFEB-dependent manner. Collectively, EA can effectively suppress ferroptosis and ferritinophagy, induce TFEB-dependent lysosomal biogenesis, and inhibit lysosomal membrane permeabilization (LMP)-mediated lysosome iron efflux into a malignant circuit in CIR.
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