SSD: Targeting inflammasome and oxidative stress as a therapeutic strategy in inflammatory diseases

This study systematically investigated the therapeutic efficacy of SSD, a stigmasterol derivative with H₂S-releasing properties, against NLRP3 inflammasome activation using both in vitro and in vivo models.

Male FPS KM mice were utilized to establish acute liver injury and myocardial fibrosis models. THP-1 and J774A.1 cell lines served for in vitro analyses, while zebrafish models were employed to assess oxidative stress responses.

SSD was administered through oral gavage at 25-100 mg/kg (acute liver injury) or 15-60 mg/kg/day (cardiac fibrosis models, daily for 7 days). In vitro experiments applied SSD at concentrations ranging from 2 to 8 μM.

Key methodologies encompassed histopathological analysis (H&E/Masson staining), serum ALT/AST quantification, Western blot for IL-1β/caspase-1 profiling, molecular docking simulations, cellular thermal shift assays (CETSA), and ROS detection via DCFH-DA probes. Statistical analysis was performed using one-way ANOVA.

SSD treatment markedly attenuated acute hepatic injury and cardiac collagen deposition. Molecular docking simulations identified direct SSD-NLRP3 binding, a finding corroborated by CETSA. SSD dose-dependently suppressed NLRP3-associated protein expression and gene transcription across hepatic, cardiac, and macrophage models, while significantly mitigating oxidative stress in both cellular and organismal systems.

SSD dually inhibits NLRP3 via direct targeting and oxidative stress modulation, demonstrating therapeutic potential for NLRP3-driven inflammation.