Remazolam reduces the activation of hepatic stellate cells by inactivating NOD-like receptor family pyrin domain containing 3/caspase-1/gasdermin D pathway

Liver fibrosis (LF) is a progressive pathological process driven by hepatic stellate cell (HSC) activation. This study aimed to explore the effect of remazolam (RE), a benzodiazepine anesthetic, on HSC activation and its underlying mechanisms. LX-2 cells were stimulated with lipopolysaccharide (LPS) and treated with various RE concentrations. Cell viability was assessed by CCK-8 assay, and the expression of fibronectin, collagen I, and ?-SMA was analyzed via western blot. Pyroptosis was evaluated by morphological observation, LDH release, ELISA for IL-1?/IL-18, and protein expression in the NLRP3/Caspase-1/GSDMD pathway. Rescue experiments were performed by overexpressing NLRP3. RE dose-dependently reduced LX-2 cell viability and downregulated fibronectin, collagen I, and ?-SMA expression. RE also significantly inhibited pyroptosis, evidenced by reduced LDH release, decreased IL-1?/IL-18 levels, and suppression of NLRP3, ASC, cleaved caspase-1, GSDMD-N, and IL-1?. Overexpression of NLRP3 reversed the inhibitory effects of RE on pyroptosis and HSC activation. These findings indicate that RE alleviates LF by inhibiting HSC activation and pyroptosis through suppression of the NLRP3/Caspase-1/GSDMD pathway, highlighting its potential as a therapeutic agent for LF.