Fig. 5: Methylcobalamin protects against GSDME-initiated liver failure.

5 dpf zebrafish larvaes were pretreated with 20 μM MeCbl before immersed in 400 μM DCA for 48 h. a–c SYTOX fluorescence probe and immunoblotting analysis were used to investigate the protective effects of MeCbl on DCA-induced systemic and liver injury (liver region was indicated with red or yellow circle, scale bar=250 μm) in zebrafish, n = 6. d Survival analysis of zebrafish after DCA immersion, n = 60. e Sequence alignment of GSDME from human, mouse and zebrafish. In addition, mice were performed with bile duct ligation (BDL) surgery to induce GSDME-mediated acute liver failure. MeCbl (10 mg/kg), DMF (50 mg/kg) were given intragastrically to mice daily before euthanized. f–j Serum ALT, AST, IL-1α, IL-1β, HMGB1 levels in WT and Gsdme^-/- mice, n = 6. k Representative immunoblots of caspase-3 and GSDME in the livers of the mice. l Survival analysis of WT and Gsdme^-/- mice after BDL and treated as indicated, n = 10. m, n Representative H&E sections of the liver (m) (scale bar = 50 μm). Damaged area was traced with black dash line and analyzed using Image J software (n, n = 6 representative section of six independent mice samples). Data are mean ± SEM of at least two independent experiments. Each point is an individual zebrafish larvae or mouse, number is indicated, n = 6-60 per group. Blots are representative data of six independent samples (c, k). Analysis was done using two-tailed unpaired Student’s t test (a) or two-way ANOVA (f–j, n). Log-rank (Mantel-Cox) test was used for survival analysis (d, l). p < 0.05 is considered significant to indicated group and p values are provided in graphs, n.s=nonsignificant. Source data are provided as Source Data file.