Journal: Redox Biology
Article Title: Epithelial redox stress programs macrophage immunometabolism through a ZNF24-MIF–NF–κB pathway in chronic nonbacterial prostatitis
doi: 10.1016/j.redox.2026.104042
Figure Lengend Snippet: Epithelial ROS-ZNF24 axis drives MIF transcription and promotes CD74-dependent M1 macrophage polarization. (A–E) CD74 knockdown attenuates MIF-induced M1 macrophage polarization, as assessed by CD86 and iNOS expression (A–B), proinflammatory cytokine secretion (C), and flow cytometry analysis of F4/80 + CD86 + macrophages (D–E). (F–H) LPS stimulation induces MIF mRNA expression (F), protein expression (G), and secretion (H) in RWPE-1 prostate epithelial cells. (I–K) Transwell co-culture system showing that LPS-stimulated prostate epithelial cells promote M1 polarization and cytokine secretion in iBMDMs, which is suppressed by the MIF inhibitor ISO-1. (L–M) Increased epithelial oxidative stress in EAP mice and LPS-stimulated RWPE-1 cells, indicated by 8-OHdG staining (L) and intracellular ROS levels (M), respectively; NAC effectively reduces ROS accumulation. (N–O) ROS scavenging with NAC suppresses epithelial MIF expression and attenuates M1 marker expression in co-cultured macrophages. (P–T) ZNF24 is induced by epithelial ROS and directly regulates MIF transcription, as shown by ZNF24 expression (P), ZNF24 knockdown (Q), predicted ZNF24 binding motifs in the MIF promoter (R), and ChIP assays demonstrating enhanced ZNF24 binding upon LPS stimulation (S–T). Data are presented as mean ± SD. ns, not significant; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. Abbreviations: iBMDMs, immortalized bone marrow-derived macrophages; RWPE-1, human prostate epithelial cell line; siCD74, small interfering RNA targeting CD74; LPS, lipopolysaccharide; NAC, N-acetylcysteine; ROS, reactive oxygen species.
Article Snippet: The iBMDM cell line (a murine immortalized macrophage line) was kindly provided by academician Feng Shao, and the human prostate epithelial cell line RWPE-1 (Cat. No. CL-0200, Procell) was obtained from Procell. iBMDM cells were cultured in high-glucose DMEM containing 10 % FBS and 1 % penicillin-streptomycin and passaged as needed.
Techniques: Knockdown, Expressing, Flow Cytometry, Co-Culture Assay, Staining, Marker, Cell Culture, Binding Assay, Derivative Assay, Small Interfering RNA
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