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KE administration improves <t>liver</t> injury in LPS-induced septic <t>mice.</t> Nine-week-old male C57BL/6 mice were orally administered 3 mg/g KE per day for 3 days, followed by intraperitoneal injection of 5 mg/kg LPS for 24 h. (A) The circulating β-OHB levels were measured (n = 6). (B) The liver sections of mice from each group were stained with H&E, Scale bar = 50 μm, (n = 3). (C,D) Measurement of the serum ALT and AST levels by colorimetry (n = 6). (E) Immunoblotting analysis of the Kbhb modification in the hepatic <t>tissues</t> (n = 3). (F,G) After immunoprecipitation using anti-SOD2 antibodies, immunoblotting analysis of the SOD2 protein expression and its Kbhb levels in hepatic tissues was performed (n = 3). (H) Measurement of the hepatic enzymatic activity of SOD2 (n = 5). (I – L) Immunoblotting analysis of the hepatic NLRP3, ASC and cleaved-caspase 1 protein expression (n = 3). (M) The IL-1β levels in the hepatic tissues measured by ELISA (n = 6). (N,O) Immunoblotting analysis of Bax and Bcl-2 protein expression in the hepatic tissues (n = 3). ∗ p < 0.05, ∗∗ p < 0.01.
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KE administration improves liver injury in LPS-induced septic mice. Nine-week-old male C57BL/6 mice were orally administered 3 mg/g KE per day for 3 days, followed by intraperitoneal injection of 5 mg/kg LPS for 24 h. (A) The circulating β-OHB levels were measured (n = 6). (B) The liver sections of mice from each group were stained with H&E, Scale bar = 50 μm, (n = 3). (C,D) Measurement of the serum ALT and AST levels by colorimetry (n = 6). (E) Immunoblotting analysis of the Kbhb modification in the hepatic tissues (n = 3). (F,G) After immunoprecipitation using anti-SOD2 antibodies, immunoblotting analysis of the SOD2 protein expression and its Kbhb levels in hepatic tissues was performed (n = 3). (H) Measurement of the hepatic enzymatic activity of SOD2 (n = 5). (I – L) Immunoblotting analysis of the hepatic NLRP3, ASC and cleaved-caspase 1 protein expression (n = 3). (M) The IL-1β levels in the hepatic tissues measured by ELISA (n = 6). (N,O) Immunoblotting analysis of Bax and Bcl-2 protein expression in the hepatic tissues (n = 3). ∗ p < 0.05, ∗∗ p < 0.01.

Journal: Redox Biology

Article Title: β-Hydroxybutyrate ameliorates lipopolysaccharide-induced liver injury through β-hydroxybutyrylation of the SOD2 protein in mice

doi: 10.1016/j.redox.2025.103949

Figure Lengend Snippet: KE administration improves liver injury in LPS-induced septic mice. Nine-week-old male C57BL/6 mice were orally administered 3 mg/g KE per day for 3 days, followed by intraperitoneal injection of 5 mg/kg LPS for 24 h. (A) The circulating β-OHB levels were measured (n = 6). (B) The liver sections of mice from each group were stained with H&E, Scale bar = 50 μm, (n = 3). (C,D) Measurement of the serum ALT and AST levels by colorimetry (n = 6). (E) Immunoblotting analysis of the Kbhb modification in the hepatic tissues (n = 3). (F,G) After immunoprecipitation using anti-SOD2 antibodies, immunoblotting analysis of the SOD2 protein expression and its Kbhb levels in hepatic tissues was performed (n = 3). (H) Measurement of the hepatic enzymatic activity of SOD2 (n = 5). (I – L) Immunoblotting analysis of the hepatic NLRP3, ASC and cleaved-caspase 1 protein expression (n = 3). (M) The IL-1β levels in the hepatic tissues measured by ELISA (n = 6). (N,O) Immunoblotting analysis of Bax and Bcl-2 protein expression in the hepatic tissues (n = 3). ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Interleukin-1β (IL-1β) levels in the supernatants of J774A.1 cells and in mouse liver tissue homogenates were analyzed using a cytokine-specific ELISA kit (#RK00006, Abclonal).

Techniques: Injection, Staining, Colorimetric Assay, Western Blot, Modification, Immunoprecipitation, Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay

The SOD2-K68R mutant attenuates the protective effects of KE on LPS-induced septic liver injury in mice. Nine-week-old male C57BL/6 mice were transduced with AAV8-SOD2 or AAV8-SOD2-K68R vectors via tail vein injection. After 2 weeks of injection, the mice were subjected to KE + LPS treatment. (A) The circulating β-OHB levels were measured (n = 6). (B) The liver sections of mice from each group were stained with H&E, Scale bar = 50 μm, (n = 3). (C,D) Measurement of the serum ALT and AST levels by colorimetry (n = 6). (E,F) After immunoprecipitation using anti-Flag beads, immunoblotting analysis of the Flag-tagged protein and its Kbhb levels in hepatic tissues was performed (n = 3). (G,H) Immunoblotting analysis of the SOD2 protein expression in the hepatic tissues (n = 3). (I) Measurement of the hepatic enzymatic activity of SOD2 (n = 5). (J – M) Immunoblotting analysis of the hepatic NLRP3, ASC and cleaved-caspase 1 protein expression (n = 3). (N) The IL-1β levels in the hepatic tissues measured by ELISA (n = 6). (O,P) Immunoblotting analysis of Bax and Bcl-2 protein expression in the hepatic tissues (n = 3). ∗ p < 0.05, ∗∗ p < 0.01.

Journal: Redox Biology

Article Title: β-Hydroxybutyrate ameliorates lipopolysaccharide-induced liver injury through β-hydroxybutyrylation of the SOD2 protein in mice

doi: 10.1016/j.redox.2025.103949

Figure Lengend Snippet: The SOD2-K68R mutant attenuates the protective effects of KE on LPS-induced septic liver injury in mice. Nine-week-old male C57BL/6 mice were transduced with AAV8-SOD2 or AAV8-SOD2-K68R vectors via tail vein injection. After 2 weeks of injection, the mice were subjected to KE + LPS treatment. (A) The circulating β-OHB levels were measured (n = 6). (B) The liver sections of mice from each group were stained with H&E, Scale bar = 50 μm, (n = 3). (C,D) Measurement of the serum ALT and AST levels by colorimetry (n = 6). (E,F) After immunoprecipitation using anti-Flag beads, immunoblotting analysis of the Flag-tagged protein and its Kbhb levels in hepatic tissues was performed (n = 3). (G,H) Immunoblotting analysis of the SOD2 protein expression in the hepatic tissues (n = 3). (I) Measurement of the hepatic enzymatic activity of SOD2 (n = 5). (J – M) Immunoblotting analysis of the hepatic NLRP3, ASC and cleaved-caspase 1 protein expression (n = 3). (N) The IL-1β levels in the hepatic tissues measured by ELISA (n = 6). (O,P) Immunoblotting analysis of Bax and Bcl-2 protein expression in the hepatic tissues (n = 3). ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Interleukin-1β (IL-1β) levels in the supernatants of J774A.1 cells and in mouse liver tissue homogenates were analyzed using a cytokine-specific ELISA kit (#RK00006, Abclonal).

Techniques: Mutagenesis, Transduction, Injection, Staining, Colorimetric Assay, Immunoprecipitation, Western Blot, Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay