Pharmacological Targeting of CXCR4 Attenuates Sepsis-Induced Intestinal Injury by Suppressing NLRP3/GSDMD-Mediated Pyroptosis.

Sepsis-induced intestinal barrier dysfunction is a critical driver of multiple organ failure and associated mortality. Although pyroptosis contributes to sepsis pathogenesis, the upstream receptors that trigger this process and constitute viable therapeutic targets remain elusive. Here, we identify C-X-C chemokine receptor type 4 (CXCR4) as a key upstream regulator of intestinal epithelial pyroptosis during septic injury. In a murine cecal ligation and puncture (CLP) model, sepsis robustly upregulated CXCR4 expression, which subsequently induced NLRP3 upregulation, caspase-1 activation, and generation of the gasdermin D N-terminal fragment (GSDMD-NT). Pharmacological inhibition of CXCR4 with the clinically approved antagonist AMD3100 (Plerixafor) significantly attenuated intestinal injury, restored the expression of tight junction proteins (ZO-1 and occludin), and suppressed pyroptotic markers. Conversely, CXCR4 agonism with NUCC-390 exacerbated intestinal damage. Mechanistically, CXCR4 activation enhanced NF-κB p65 phosphorylation, thereby propelling NLRP3-dependent GSDMD cleavage. Genetic knockdown of GSDMD abolished NUCC-390-induced pyroptosis and barrier impairment without affecting NF-κB activation, confirming GSDMD as the essential downstream executor. Furthermore, pharmacological inhibition of NF-κB abrogated the exacerbation of intestinal injury, barrier dysfunction, and pyroptosis induced by CXCR4 activation, definitively establishing NF-κB as a necessary signaling mediator within this CXCR4-driven pathway. These findings establish the CXCR4/NF-κB/NLRP3/GSDMD axis as a novel and therapeutically targetable pathway in septic intestinal injury. Importantly, we demonstrate that the FDA-approved CXCR4 antagonist AMD3100 represents a promising preclinical therapeutic candidate, exhibiting dual anti-inflammatory and anti-pyroptotic efficacy. Our work thus provides a mechanistically grounded rationale for further investigation of its repurposing potential in sepsis.