Carbon Monoxide Protects against Hepatic Ischemia/Reperfusion Injury via ROS-Dependent Akt Signaling and Inhibition of Glycogen Synthase Kinase 3β
Figure 2
Inhibition of GSK3β by CO inhalation ameliorates liver I/R injury. Mice were sham-operated or subjected to 90 minutes hepatic warm ischemia followed by 1 hour reperfusion. Recipients were treated with air or CO gas (250 ppm) inhalation. (a) Liver samples, harvested 1 hour later, were subjected to Western blot analysis of phospho (p)-GS (GS 641), p-GSK3β (S9), p-JNK, and p-p38. β-Actin was used as an internal control. , . (b) Immunohistochemical staining of GSK3 inhibition. Mice were sacrificed, liver tissues were harvested, and the tissue slices were processed for formalin-fixed paraffin embedding. Hematoxylin counterstaining after 3-amino-9-ethylcarbazole-based immunohistochemical staining was used to detect GS 641 ((A)–(C), red/brown) and phosphorylation of GSK3β S9 ((D)–(F), red/brown). ((c) and (d)) Quantitative RT-PCR-assisted detection of TNF-α, IL-6, CXCL10, and IL-10 gene expression at 1 hour or 6 hours in liver tissue. Data were normalized to 18S gene expression. Data shown represent the mean ± S.D. ( = 4-5/group), .
