CXCR4 inhibition alleviates prostatic inflammation and pelvic pain via suppressing Th17 cell differentiation and oxidative stress in EAP mice

CXCR4抑制剂通过抑制Th17细胞分化和氧化应激,减轻EAP小鼠的前列腺炎症和盆腔疼痛。

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Abstract

Background: Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) represents a prevalent urological disorder characterized by urinary symptoms, persistent pelvic or perineal discomfort accompanied by intraprostatic leukocyte infiltration. The C-X-C chemokine receptor type 4 (CXCR4) is critically involved in mediating inflammatory responses. Nevertheless, the specific involvement of CXCR4 in the immunoinflammatory mechanisms underlying CP/CPPS pathogenesis remains poorly characterized. Methods: The therapeutic efficacy of AMD3100, a CXCR4 antagonist, in CP/CPPS was evaluated in a murine model of experimental autoimmune prostatitis (EAP). The progression of EAP and T helper 17 (Th17) cell-mediated immune responses following AMD3100 intervention was assessed via HE staining, immunohistochemistry, immunofluorescence, quantitative polymerase chain reaction (qPCR), and flow cytometry. To unravel mechanistic insights into the role of CXCR4 in regulating Th17 cell differentiation, RNA sequencing, qPCR, and western blotting validation were conducted. In addition, histological staining, measurements of reactive oxygen species (ROS) and peroxidation markers, and co-culture assays were employed to assess the antioxidative effects of AMD3100 in prostate epithelial cells. Results: AMD3100 significantly alleviated a series of symptoms of prostatitis in EAP mice. Meanwhile, inhibition of CXCR4 by AMD3100 could significantly decrease the proportion of Th17 cells and downregulate the elevated expression of both pro-inflammatory and Th17-associated cytokines in these mice. However, administration of IL-17A partially reversed the therapeutic effects of AMD3100, elevating oxidative stress biomarkers and promoting the apoptosis of prostate epithelial cells. Mechanistically, CXCR4 inhibition suppresses NF-κB activation, thereby inhibiting Th17 cell differentiation. Furthermore, integrated findings from both in vitro and in vivo studies demonstrated that aberrant NF-κB activation not only counteracted AMD3100-mediated suppression of Th17 cell differentiation but also exacerbated prostatic epithelial cell damage through amplified inflammatory responses, oxidative stress, and apoptosis. Conclusions: CXCR4 presents a promising therapeutic target for CP/CPPS. Pharmacological blockade of CXCR4 with AMD3100 inhibits Th17 cell differentiation, consequently mitigating inflammatory infiltration and oxidative tissue injury in CP/CPPS.

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