Fig. 1

Chemical structure of rebamipide. A Overview of the experimental design. B Protective effects of rebamipide on α-synuclein + MPP⁺-induced cytotoxicity in BV2 microglia cells. C BV2 microglia cells were treated with rebamipide for 1 h and then stimulated with α-synuclein + MPP⁺ for an additional 11 h. Lactate dehydrogenase (LDH) (C), TNF-α (D), IL-6 (E), IL-1ß (F, G, and I), pro-IL-1ß (H), and IL-18 (G) levels were evaluated. Inhibitory effects of rebamipide on NLRP3 (J), ASC (K), pro-caspase-1 (L), and p20 (M) in BV2 microglia cells were measured using ELISA. Molecular docking illustrating the binding interactions of rebamipide with the NLRP3-NEK7 complex and NLRP3 alone (N). The left panels depict 3D docking models, whereas the right panels represent 2D interaction diagrams highlighting key binding residues. Hydrogen bonds are indicated by purple arrows, halogen bonds by yellow arrows, and salt bridges by red and blue lines. Rebamipide interacts with ASN978, TYR1009, and PRO1034 of NLRP3 and LYS163 of NEK7, suggesting a role in disrupting the NLRP3-NEK7 interaction and modulating inflammasome activation. Surface plasmon resonance (SPR) sensorgrams demonstrating the real-time binding kinetics of rebamipide to NLRP3-NEK7 complex (O) and NLRP3 alone (P). Rebamipide exhibited dose-dependent binding, with a higher binding affinity for NLRP3-NEK7 complex than NLRP3 alone. Overview of the experimental design (Q). Inhibitory effects of rebamipide on NLRP3-induced upregulation of IL-1ß (R) and IL-18 (S) in BV2 microglia cells. BV2 microglia cells were treated with rebamipide for 1 h and then stimulated with MSU, nigericine, ATP, and hemozoin for an additional 11 h. IL-1ß and IL-18 levels were evaluated (T-X). Data are presented as mean ± standard error of mean (SEM). * p < 0.05, ** p < 0.01, ***p < 0.001, compared with the control group; # p < 0.05, ## p < 0.01, ### p < 0.001, compared with the α-synuclein + MPP⁺ or NLRP3 inducers-treated groups