Histone H4 lysine 12 lactylation-driven ZIP14 expression is involved in electroacupuncture therapy against cerebral ischemia/reperfusion injury in mice

• EA decreased lactate levels and histone H4 lysine 12 lactylation (H4K12la) in neurons following cerebral ischemia/reperfusion (I/R). • Decreased neuronal PKM2 expression mediates H4K12la upregulation in response to EA. • H4K12la promotes the transcription Zip14 and triggers ferroptosis following cerebral I/R. • EA exerts neuroprotective effects against ferroptosis via inhibition of H4K12la-dependent ZIP14 expression. • EA decreases arterial lactate level after reperfusion in stroke patients. Electroacupuncture (EA) has been demonstrated as an effective therapeutic intervention for cerebral ischemia–reperfusion injury (CIRI); however, the fundamental processes underlying EA therapy remain largely elusive, which hinders the optimization and broader clinical application of EA. It has been reported that during CIRI, cellular metabolism undergoes a shift from oxidative phosphorylation to glycolysis, giving rise to an accumulation of lactate, but whether and how lactate is involved in CIRI and EA therapy is not fully understood. To explore the role of lactate in EA therapy against CIRI and the underlying mechanisms. Neurological outcome evaluations and TTC staining were performed to assess CIRI in mice. Western blotting and immunofluorescence were used to detect histone lactylation, and genes regulated by histone lactylation were identified by CUT&Tag, ATAC-seq and RNA-seq, followed by investigation of the role(s) of an iron transporter encoding gene Zip14 in EA therapy. Arterial lactate levels in stroke patients were assessed 30 min after recanalization using arterial blood gas analysis. Lactate markedly enhances histone H4 lysine 12 lactylation (H4K12la) in neurons, concomitantly upregulating PKM2 expression—a pivotal regulator of lactate production. EA protects neurons in the ischemic penumbra and improves neurological outcomes following CIRI by suppressing PKM2-mediated H4K12la. Furthermore, CIRI elevates H4K12la enrichment at Zip14 , which facilitates chromatin accessibility, activates Zip14 transcription, and ultimately triggers ferroptosis. EA treatment attenuates ferroptosis and mitigates CIRI by decreasing ZIP14 expression. EA significantly reduces arterial lactate levels after recanalization in patients with ischemic stroke. This study uncovered a previously unrecognized mechanism by which lactate is involved in EA therapy against CIRI, highlighting H4K12la-dependent ZIP14 expression as a potential therapeutic target for CIRI management and EA optimization.