Carbon Monoxide Protects against Hepatic Ischemia/Reperfusion Injury via ROS-Dependent Akt Signaling and Inhibition of Glycogen Synthase Kinase 3β
Figure 1
Pretreatment of mice with CO gas inhalation ameliorates liver I/R injury via AKT-GSK3β activation. Mice were subjected to 90 minutes liver warm ischemia, followed by 6 h reperfusion. (a) Hepatocellular function was evaluated by sALT (IU/L). (b) Representative liver histology of ischemic liver lobes. (c) Liver neutrophil accumulation, assessed by MPO activity. Data represent the mean ± standard deviation (SD) ( = 4–6 samples/group). . (d) Hepatic HMGB1 expression in liver tissue was assessed by Western blot analysis at 1 h and 6 h of reperfusion. Total cell lysates were analyzed for HMGB1 and β-actin protein levels by Western blot analysis. (e) Western-blot analysis of phospho (p)-GSK3β (Ser 9), p-GS (Ser641), and p-Akt in HepG2 cells after treatment with CORM2 (50 μM) at the indicated times. (f) RAW264.7 cells were stimulated with 10 ng/mL of LPS for 30 minutes in the absence or presence of CORM2 and the ROS scavenger, N-acetyl-cysteine (NAC). Total cell lysates were analyzed for phosphorylated GS, GSK3β, and Akt as well as total GS, GSK3β, Akt, and β-actin protein levels by Western immunoblot analysis. (g) Mice were subjected to 90 minutes of liver warm ischemia, followed by 1 h or 6 h reperfusion. Liver tissue was analyzed by Western blotting of p-Akt and total Akt. β-Actin served as the standard.
