Gadolinium chloride alleviates partial warm ischemia-reperfusion injury of liver by suppressing kupffer cell function in mice

Abstract

Objective To investigate the effect and mechanism of gadolinium chloride alleviates partial warm ischemia-reperfusion injury of liver by suppressing kupffer cell function in mice. Methods Forty-eight male C57BL/6 mice were randomly divided into sham-operated group (Sham), Hepatic ischemia-reperfusion injury (HIRI) group, gdolinium chloride groups (Gd+ IRI), and cobalt protoporphyrin (CoPP+ IRI) group (n=12). The rats were injected with the same dose of normal saline in Sham group and IRI group. Gd+ IRI group were intraperitoneally injected with GdCl3 (20 mg/kg) 24 hours before operation. At the same time, CoPP+ IRI group were intraperitoneally injected with CoPP (5 mg/kg). Except for Sham group, an 70% volume HIRI model was established by means of 60 minutes ischemia and then 6 hours reperfusion in the other groups. The levels of serum aspartate transaminase (AST) and alanine aminotransferase (ALT) in each group were compared. Enzyme-linked immunosorbent assay were used to test the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in the liver tissue. The level of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, heme oxygenase-1 (HO-1), and protective factor IL-10 mRNA level were detected by real time quantitative polymerase Chain Reaction (RT-PCR). Immunohistochemical staining was used to measure Fas, FasL protein expression level of extrahepatic biliary epithelial cells. Results Respectively, the serum ALT level in the HIRI, Gd+ IRI and CoPP+ IRI groups was (1 236.7±62.4), (638.9±50.9), (740.3±53.1) U/L, and the AST level was (1 107.8±68.3), (568.5±42.7), (679.1±49.9) U/L, the serum level of ALT and AST in Gd+ IRI and CoPP+ IRI groups was lower than IRI group (t=6.721, P<0.01; t=4.732, P<0.05; t=5.984, P<0.01; t=2.691, P<0.05). Compared with IRI group, the activity of SOD and GSH-Px was promoted (t=7.301, P<0.01; t=1.937, P<0.05), whereas the levels of MDA was reduced (t=9.519, P<0.01; t=8.635, P<0.01). Simultaneously, the TNF-α, IL-1β and IL-6 mRNA level in Gd+ IRI and CoPP+ IRI groups were decreased (t=6.721, P<0.01; t=2.316, P<0.05), and the levels of IL-10 and HO-1 mRNA were increased (t=2.973, P<0.05; t=7.921, P<0.01). After hepatic artery reflow 6 h, the Fas expression levels on bile ducts epithelial cells in Gd+ IRI and CoPP+ IRI groups respectively was 22.4%, 29.7%, which was lower than IRI group (38.9%) (χ2=6.561, P<0.01; χ2=2.547, P<0.05). The FasL expression levels in Gd+ IRI and CoPP+ IRI groups respectively was 21.2%, 25.1%, which was lower than IRI group (36.5%) (χ2=5.982, P<0.05; χ2=3.188, P<0.05). Conclusion Gdolinium chloride shows an protective function aganist liver ischemia-reperfusion iniury in mice by Oxidative stress, inhibiting inflammation and cell apoptosis. The up-regulation of HO-1 by gdolinium chloride may be the possible mechanism. The up-regulation of HO-1 and inhibiting the Fas/FasL protein expression by gdolinium chloride may be the possible mechanism. Key words: Gdolinium chloride; Liver; Ischemia-reperfusion injury; Heme oxygenase-l