ADK expression reduction promotes liver IRI. (A) RNA-seq revealed the mRNA level of ADK after IRI in mouse liver tissue. (B) ADK mRNA level in mouse liver IRI transcriptome dataset GSE93034 was shown. (C and D) Protein levels of ADK after IRI in mouse livers were detected by immunoblotting and immunohistochemistry. Scale bar: 200 μm (D). (E and F) ADK protein levels were detected in donor livers before reperfusion or after reperfusion by immunoblotting (E). Comparison of protein expression levels of ADK before and after reperfusion (F). n = 24 for donor livers (F). (G–I)Adkf/f, Adkf/f;Alb-Cre, or Adkf/f;Alb-Cre;Ripk1D138N/D138N mice underwent 1-h ischemia/6-h reperfusion operation at the age of 5 wk. The levels of p-RIPK1(S166), CC8, CC3, and ADK in isolated hepatocytes were determined by immunoblotting (G). Liver H&E staining (H) and serum ALT/AST detection (I) were performed. Scale bar: 100 μm (H). (J–L) Vehicle or 5-ITu (20 mg/kg) was injected intraperitoneally to WT mice at the age of 8 wk. 1 h after the injection, the mice underwent 1-h ischemia/6-h reperfusion operation. The levels of p-RIPK1(S166), CC8, CC3, and ADK were determined by immunoblotting (J). Liver H&E staining (K) and serum ALT/AST detection (L) were performed. Scale bar: 100 μm (K). (M–O) AAV8 that was expressing Adk-L or control was injected to WT mice through tail vein. 1 mo after the injection, the mice underwent 1-h ischemia/6-h reperfusion operation. The levels of p-RIPK1(S166), CC8, CC3, and ADK were determined by immunoblotting (M). Liver H&E staining (N) and serum ALT/AST detection (O) were performed. Scale bar: 100 μm (N). Data are represented as the mean ± SD (A, B, H, I, K, L, N, and O). Data are representative of n = 3 independent experiments (A–O). Statistical significance was determined using unpaired two-tailed Student’s t test (A and B), paired two-tailed Student’s t test (F), or two-way ANOVA with post hoc Bonferroni’s test (H, J, K, L, N, and O). ***P < 0.001. Source data are available for this figure: SourceData F8.
Figure 8.

ADK expression reduction promotes liver IRI. (A) RNA-seq revealed the mRNA level of ADK after IRI in mouse liver tissue. (B) ADK mRNA level in mouse liver IRI transcriptome dataset GSE93034 was shown. (C and D) Protein levels of ADK after IRI in mouse livers were detected by immunoblotting and immunohistochemistry. Scale bar: 200 μm (D). (E and F) ADK protein levels were detected in donor livers before reperfusion or after reperfusion by immunoblotting (E). Comparison of protein expression levels of ADK before and after reperfusion (F). n = 24 for donor livers (F). (G–I)Adkf/f, Adkf/f;Alb-Cre, or Adkf/f;Alb-Cre;Ripk1D138N/D138N mice underwent 1-h ischemia/6-h reperfusion operation at the age of 5 wk. The levels of p-RIPK1(S166), CC8, CC3, and ADK in isolated hepatocytes were determined by immunoblotting (G). Liver H&E staining (H) and serum ALT/AST detection (I) were performed. Scale bar: 100 μm (H). (J–L) Vehicle or 5-ITu (20 mg/kg) was injected intraperitoneally to WT mice at the age of 8 wk. 1 h after the injection, the mice underwent 1-h ischemia/6-h reperfusion operation. The levels of p-RIPK1(S166), CC8, CC3, and ADK were determined by immunoblotting (J). Liver H&E staining (K) and serum ALT/AST detection (L) were performed. Scale bar: 100 μm (K). (M–O) AAV8 that was expressing Adk-L or control was injected to WT mice through tail vein. 1 mo after the injection, the mice underwent 1-h ischemia/6-h reperfusion operation. The levels of p-RIPK1(S166), CC8, CC3, and ADK were determined by immunoblotting (M). Liver H&E staining (N) and serum ALT/AST detection (O) were performed. Scale bar: 100 μm (N). Data are represented as the mean ± SD (A, B, H, I, K, L, N, and O). Data are representative of n = 3 independent experiments (A–O). Statistical significance was determined using unpaired two-tailed Student’s t test (A and B), paired two-tailed Student’s t test (F), or two-way ANOVA with post hoc Bonferroni’s test (H, J, K, L, N, and O). ***P < 0.001. Source data are available for this figure: SourceData F8.

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