AR blockade accelerates healing and directly inhibits macrophage TNF-α expression. (a) Flutamide treatment significantly accelerates healing, as illustrated by reduced wound cross-sectional areas at day 3 after wounding (*P < 0.05) and by histological analysis (b) of day 3 wounds (arrows demarcate wound edge). (c) Reduced TNF-α expression at day 3 after wounding following flutamide treatment (F, oral flutamide; C, vehicle control; NS, normal skin from control animal). HPRT was used as the housekeeping gene. Bottom panel represents EMSA illustrating increased NF-κB binding activity in day 3 wounds (control) compared with normal skin; binding is markedly reduced in the day 3 wounds of flutamide-treated animals. (d) TGF-β1 (TGF)acted as a potent chemoattractant for murine peritoneal macrophages (C, control medium; *P < 0.05). Testosterone (Testo) had no effect on chemotaxis when used in conjunction with TGF-β1 or alone as a chemoattractant, nor when the cells were pretreated with testosterone. (e) LPS-induced TNF-α mRNA expression by murine macrophages. C, control; +, LPS. Testosterone markedly increased TNF-α expression both basally (T) and when cells were LPS-activated (T+). Flutamide treatment significantly reduced testosterone-induced TNF-α expression levels. Wound tissue was pooled from four mice, and the gel shown is representative of three experiments.