Cis-resveratrol blocks crystal-induced NLRP3 inflammasome activation via the TRPV4-Ca²⁺-phagocytosis-ROS axis

s Background Inflammation is a vital biological defense mechanism, yet uncontrolled or prolonged inflammation can cause tissue damage. The NLRP3 inflammasome, a central component of innate immunity, drives inflammation by activating caspase-1 and promoting the release of cytokines like IL-1β. Monosodium urate (MSU) crystals are potent NLRP3 inflammasome activators implicated in diseases such as gout. While resveratrol, a natural polyphenol, exhibits anti-inflammatory properties, the specific mechanisms by which its cis-isomer (Cis-Resv) inhibits NLRP3 inflammasome activation remain poorly understood. Purpose This study aims to investigate the inhibitory effect of Cis-Resv on MSU-induced NLRP3 inflammasome activation and elucidate the underlying molecular mechanisms. Methods Bone marrow-derived macrophages (BMDMs) from C57BL/6J mice were stimulated with MSU crystals. The effects of Cis-Resv on IL-1β secretion, caspase-1 activation, and ASC oligomerization were evaluated using ELISA, Western blot, and immunoprecipitation, respectively. Transcriptomic analysis was employed to identify potential signaling pathways involved. The in vivo efficacy of Cis-Resv was assessed in MSU-induced acute gouty arthritis and air-pouch models by measuring joint swelling, inflammatory cytokines levels, and histopathological changes. Results Cis-Resv significantly inhibited MSU-induced NLRP3 inflammasome activation in BMDMs, reducing caspase-1 activation, IL-1β secretion, andassociated pyroptosis. Furthermore, Cis-Resv impeded ASC oligomerization, a critical step in inflammasome assembly. Transcriptomic analysis and subsequent functional validation revealed that Cis-Resv exerts these effects primarily through the TRPV4-Ca²⁺-phagocytosis-ROS signaling axis. Specifically, Cis-Resv inhibited macrophage TRPV4 channel activity, leading to decreased Ca²⁺ influx, impaired MSU crystal phagocytosis, and attenuated reactive oxygen species (ROS) accumulation. This reduction in ROS modulated the ROS-TXNIP pathway, diminishing the interaction between TXNIP and NLRP3, thereby suppressing inflammasome activation. In vivo , Cis-Resv effectively alleviated MSU-induced joint swelling, lowered IL-1β and TNF-α levels, and ameliorated histopathological damage in both gouty arthritis and air-pouch models. Conclusion Cis-Resv suppresses MSU-induced NLRP3 inflammasome activation by targeting the TRPV4-Ca²⁺-phagocytosis-ROS axis. This study identifies a novel mechanism and suggests Cis-Resv as a potential therapeutic agent for NLRP3-driven inflammatory conditions like gout.