ADK expression reduction aggravates liver IRI. (A and B) Primary hepatocytes from WT mice were treated with CHX (1 μM)/TNFα (10 ng/ml) for the indicated time (A) or 12 h (B). Cell death was measured by the SYTOX Green positivity assay (A). The levels of p-RIPK1(S166), CC8, and CC3 were determined by immunoblotting (B). (C–G)Ugdh^f/f, Ugdh^f/f;Alb-Cre, Ugdh^f/f;Alb-Cre;Ripk1^D138N/D138N mice were subjected to 1-h ischemia/6-h reperfusion model. Serum ALT/AST detection (C), H&E staining (D), CC3 immunostaining (E), MPO immunostaining (F), and serum TNFα and IL-6 level detection (G) were performed. Scale bar: 100 μm (D–F). (H) Protein expression level of UGDH and ADK in clinical donor liver IRI samples was analyzed with immunoblotting. (I–M)Adk^WT/f, Adk^WT/f;Alb-Cre, Adk^WT/f;Alb-Cre;Ripk1^D138N/D138N mice were fed with choline-deficient high-fat diet (60% fat, 0.1% methionine, and no added choline, A06071302; Research Diets) for 6 wk to construct the NASH model. Liver H&E staining, CC3 immunostaining, and p-RIPK1 (S166) immunostaining were performed (I), serum ALT/AST was measured (J), proinflammatory gene expression was analyzed (K), liver fibrosis was analyzed with Masson’s trichrome staining and Sirius red staining (L), and fibrotic gene expression was analyzed (M). Scale bar: 100 μm (I and L). (N) Protein expression level of UGDH and ADK in clinical NASH samples was analyzed with immunoblotting. Data are represented as the mean ± SD of n = 3 (A, C–G, and I–M). Data are representative of n = 3 independent experiments (A–N). Statistical significance was determined using two-way ANOVA with post hoc Bonferroni’s test (C–G, K, and M) or one-way ANOVA with post hoc Dunnett’s test (I, J, and L). *P < 0.05; ***P < 0.001. Source data are available for this figure: SourceData FS5.