CCR5 protects against permanent brain ischemia by suppressing neuronal ferroptosis through AMPK/ACC signaling.

Ischemic stroke triggers devastating neuronal injury cascades, and ferroptosis acts as a key mediator of post-ischemic neuronal loss. Although C-C chemokine receptor type 5 (CCR5) has been traditionally implicated in post-stroke neuroinflammation, its cell-specific role in neuronal ferroptosis remains obscure. Here, we combined single-cell transcriptomic analysis, a permanent middle cerebral artery occlusion (pMCAO) mouse model, and in vitro hypoxia-ischemia-mimicking conditions to define an endogenous neuroprotective role for neuronal CCR5. Single-cell RNA sequencing revealed a marked upregulation of the CCL-CCR signaling pathway, with CCR5 identified as a prominent hub mediating intercellular interactions. In vivo, CCR5 was significantly elevated in peri-infarct neurons from 48 h up to one month after pMCAO, rather than microglia or astrocytes, suggesting an intrinsic stress response. In vitro, hypoxia-ischemia-mimicking conditions in SH-SY5Y cells and primary cortical neurons recapitulated this compensatory yet insufficient CCR5 induction alongside HIF-1α upregulation, GPX4 depletion, and severe lipid peroxidation. Functional studies demonstrated that CCR5 knockdown or pharmacological inhibition with maraviroc (MVC) further aggravated GPX4 reduction and lipid peroxide accumulation, whereas CCR5 overexpression substantially restored anti-ferroptotic capacity. CCR5 positively regulated AMPK and ACC phosphorylation, and inhibition of AMPK completely abolished CCR5-mediated GPX4 preservation and anti-ferroptotic defense. In vivo, MVC administration further accelerated peri-infarct iron deposition, reduced neuronal survival, and significantly worsened long-term motor deficits in pMCAO mice. Collectively, these findings identify a previously unrecognized endogenous adaptive mechanism whereby ischemia-induced neuronal CCR5 limits ferroptosis through AMPK/ACC-mediated GPX4 maintenance, providing a novel therapeutic conceptualization for strengthening endogenous protective cues in ischemic stroke.