Objective To investigate the role of macrophage migration inhibitory factor(MIF)on liver inflammation in mice with a-cute liver failure(ALF).Methods Eighteen male mice were randomly divided into normal group,model group and MIF antagonist ISO-1 group.D-galactose combined with lipopolysaccharide were used to induce ALF model.The serum levels of alanine aminotrans-ferase(ALT)、aspartate aminotransferase(AST)and total bilirubin(TBIL)were detected.The pathological changes of liver were ob-served by HE staining and the apoptosis of liver cells was observed by Tunel staining.The levels of MIF,CXC chemokine receptor 2(CX-CR2)and tumor necrosis factor-α(TNF-α)in mouse liver tissues were detected by enzyme-linked immunosorbent assay(ELISA)and the expression of MIF was observed by immunofluorescence.Results Compared with the normal group,the model group showed seri-ous liver tissue structure damage,increased hepatocyte apoptosis and decreased liver function.ISO-1 reduced liver tissue damage and hepatocyte apoptosis.The expression levels of MIF,CXCR2 and TNF-α in liver tissue of mice in model group was significantly in-creased,while ISO-1 could significantly reduce the expression levels of MIF,CXCR2 and TNF-α,with statistical significance(P＜0.05).Conclusion The expression of MIF is significantly increased in ALF,and inhibition of MIF can improve liver tissue injury and the expression of inflammatory factors,indicating that MIF plays an important role in ALF.

Objective:To explore the protective effect of AGK2, a selective inhibitor of sirtuin 2 (SIRT2), on the mitochondria of L02 hepatocytes induced by thioacetamide (TAA) and its related mechanism.Methods:Human-derived hepatocyte line L02 cells were cultured in vitro. Different concentrations of SIRT2 inhibitor AGK2 were used as intervention drugs. Cell counting kit-8 (CCK8) was used to detect the effects of different concentrations of AGK2 on the activity of L02 cells, and the appropriate concentration was selected as the AGK2 intervention group. The normal group was not given any drug intervention. The model group was given 90 mmol/L TAA for modeling. Low, medium and high dose AGK2 groups were added with 1, 2 and 4 μmol/L AGK2, respectively 2 h before modeling. CCK8 was used to detect cell activity in each group. Morphological changes of cells were observed under inverted light microscope. The relative protein expression levels of isocitrate dehydrogenase (IDH1), malate dehydrogenase (MDH1), SIRT2 and fission protein 1 homologue (FIS1) were detected by Western blot. The expression of SIRT2 in cells of each group was observed by confocal laser scanning microscope. The mitochondrial membrane potential of cells in each group was observed under a fluorescence microscope. Results:When AGK2 concentration was 1, 2 and 4 μmol/L, the survival rate of cells were 98.05%, 95.76% and 91.65%, respectively, with no statistical significance compared with normal group (all P>0.05). When AGK2 concentration was 8, 16, 32, 64, 128 μmol/L, the cell survival rate was significantly decreased compared with normal group (all P<0.05). Compared with the model group, the L02 cells in low, medium and high AGK2 groups had better activity and adherence, and the floating cells were significantly reduced. The higher the concentration of AGK2, the better the cell activity and adherence, and the less floating cells. Compared with the model group, the red fluorescence of L02 cells in AGK2 group was enhanced, while the green fluorescence was weakened. The higher the AGK2 concentration was, the stronger the red fluorescence was, and the weaker the green fluorescence was. Compared with the model group, the fluorescence of SIRT2 in L02 cells of low, medium and high AGK2 groups was weakened, and the higher the concentration of AGK2, the weaker the fluorescence of SIRT2. The protein expressions of IDH1 and MDH1 in L02 cells of low, medium and high AGK2 groups were significantly higher than those of model group (all P<0.05), and were positively correlated with the concentration of AGK2 ( r=0.818, P<0.05; r=0.960, P<0.05); the protein expressions of SIRT2 and FIS1 were significantly lower than those of the model group (all P<0.05), and were negatively correlated with the concentration of AGK2 ( r=-0.992, P<0.05; r=-0.998, P<0.05). Conclusions:AGK2 can reduce the mitochondrial membrane potential stimulated by TAA in L02 cells, increase the protein expression of IDH1 and MDH1, and inhibit the protein expression of SIRT2 and FIS1 in L02 cells in a dose-dependent manner.