Epithelial redox stress programs macrophage immunometabolism through a ZNF24-MIF-NF-κB pathway in chronic nonbacterial prostatitis.

Chronic nonbacterial prostatitis (CNP) is a prevalent and refractory urogenital disorder whose immunopathogenic mechanisms remain incompletely understood. Given that redox imbalance is increasingly recognized as a critical driver of chronic inflammation, this study systematically investigated the role of epithelial redox stress in immune regulation during CNP and its underlying molecular mechanisms. By integrating plasma cytokine profiling, bulk and single-cell transcriptomic analyses, and experimental autoimmune prostatitis (EAP) models, we identified epithelial-derived macrophage migration inhibitory factor (MIF) as a central mediator driving chronic prostatic inflammation. Mechanistically, inflammatory injury induced excessive accumulation of reactive oxygen species (ROS) in epithelial cells, which in turn activated the redox-responsive transcription factor ZNF24 to bind the MIF promoter and promote its transcription. Epithelial cell-derived MIF acted in a paracrine manner on CD74-expressing macrophages. Engagement of CD74 by MIF stabilized PKM2 expression, enhanced macrophage glycolytic reprogramming, promoted PKM2 nuclear translocation, and activated NF-κB-dependent transcriptional programs, thereby driving M1 macrophage polarization and proinflammatory cytokine production. Pharmacological interventions targeting distinct key nodes of this signaling pathway-including inhibition of MIF (ISO-1), blockade of CD74 (neutralizing antibodies), stabilization of PKM2 tetramers (DASA-58), and suppression of NF-κB (JSH-23)-significantly attenuated prostatic inflammation, restored mitochondrial homeostasis, and alleviated pelvic pain in vitro or in vivo. Collectively, these findings define an epithelial ROS-ZNF24-MIF-macrophage CD74-PKM2-NF-κB signaling axis, through which coordinated enhancement of glycolytic reprogramming and inflammatory signaling promotes M1 macrophage polarization and drives the initiation and progression of CNP. Moreover, multiple redox-sensitive nodes within this pathway represent promising therapeutic targets for precision immunomodulation in CNP.