Phaseoloidin, a homogentisic acid glucoside from Entada phaseoloides, suppresses gout inflammation via NLRP3 inflammasome.

Abstract

BACKGROUND: Gout is an arthritic disorder caused by the deposition of sodium urate crystals in the joints. Phaseoloidin (PHA), a β-glucoside of homogentisic acid isolated from the seeds of the legume "Entada phaseoloides (L.) Merr.", has previously showed anti-inflammatory properties, but its effects on gout and underlying mechanisms remain unexplored. PURPOSE: This study aimed to investigate the novel therapeutic potential of PHA in gout, with a focus on its multi-target actions against NLRP3 inflammasome activation and associated inflammatory cascades-areas not yet addressed for this compound. METHOD: In vivo, arthritis models include the acute gouty arthritis model by injecting sodium urate crystals into the paws of mice, as well as the air pouch acute gout model by injecting sodium urate crystals into the subcutaneous tissue of the back of mice. In vitro, mouse peritoneal macrophages (MPM) and rat articular chondrocytes were stimulated with LPS plus sodium urate crystals, both with and without pre-treatment of PHA. Through a combination of pathological analysis, Western blot, network pharmacology analysis, immunofluorescence, immunohistochemistry, and ELISA, the anti-inflammatory activity of PHA and its therapeutic effects mediated via NLRP3 targeting were comprehensively demonstrated. RESULTS: In vivo and in vitro GA models, PHA significantly suppressed NLRP3 inflammasome activation by inhibiting caspase-1-dependent IL-1β maturation and blocking the caspase-11-GSDMD pyroptotic axis, and attenuated inflammatory cascades through reduced NETosis formatted. Notably, PHA also uniquely preserved cartilage matrix integrity by downregulating collagen-degrading enzymes. CONCLUSION: These findings reveal, for the first time, the multi-target anti-gout activity of PHA, highlighting its dual role in modulating NLRP3 inflammasome-driven inflammation and protecting joint structure. These results position PHA as a promising and innovative natural lead compound for the development of multi-mechanistic therapies against gout and related inflammatory arthritis diseases.