| Objective: To research the role of the high mobility group box protein 1(HMGB1) in drug-induced liver injury (DILI) process in Wistar rats induced by administering carbon tetrachloride ,discuss the relationship between HMGB1 and liver functions, evaluate the effects of diammonium glycyrrhizinate (DGL) and S-adenosylmethionine (SAMe) in DILI therapy.Methods: 80 wistar rats (40 male and 40 female) weighting 200±20 grams were purchased from the Expremental Animal Center of Hebei Medical University. After acclimatized for 7 days ,all rats were randomly divided into eight groups ,Group A, B, C, D, E, F, G and H,10 rats in every group.The rats in group A were given normal saline alone by intragastric administration. Groups B, C, D, E, F, G and H were separately given 5ml/kg of 50% carbon tetrachloride by subcutaneous injection in first time, and then given 2.5ml/kg of 50% carbon tetrachloride by subcutaneous injection with 2 times per week to establish drug-induced liver injury animal models. Groups C,D and E were separately given diammonium glycyrrhizinate 375mg/kg/d, ademetionine 0.833g/kg/d, diammonium glycyrrhizinate 375mg/kg/d and ademetionine 0.833g/kg/d for four weeks by intragastric administration simultaneously.Two rats dead in 4th week in group E. 50% carbon tetrachloride were given in Groups F, G and H for two weeks. And then group F was given diammonium glycyrrhizinate 375mg/kg/d, group G was given ademetionine 0.833g/kg/d, group H was given diammonium glycyrrhizinate 375mg/kg/d and ademetionine 0.833g/kg/d for two weeks by intragastric administration simultaneously.All rats in each group were fasted and sacrificed at the end of the fourth week,the blood samples were collected for measuring the liver function marker of alanine aminotransferase(ALT), aspartate amino transferase(AST),total bilirubin (TBIL),γglutamyl transferase(GGT) with automatic biochemistry analyzer.The animal liver samples were taken for observing the expression of HMGB1 in each group with immunohistochemistry method.Results:1 Liver histopathological findingsNormal liver cells, normal liver cell cord, little brown granules in cytoplasmic were showed in group A. Obvious swelling of liver cells, disordered arrangement of liver cell cord, diffuse distribution of piecemeal necrosis and steatosis,increased liver cell volume, a large brown granules in cytoplasm in group B. Swelling cells, irregular arrangement of liver cell cord, little steatosis, scattered piecemeal necrosis and brown granules in cytoplasm in group C. Swelling cells, irregular arrangement of liver cell cord, scattered piecemeal necrosis, steatosis and brown granules in cytoplasm in group D and E. Mild swelling cells, regular arrangement of liver cell cord, scattered piecemeal necrosis, steatosis and brown granules in cytoplasm were showed in group F. Mild swelling cells, regular arrangement of liver cell cord, little piecemeal necrosis and steatosis, scattered brown granules in group G. Mild swelling cells, regular arrangement of liver cell cord, no obvious piecemeal necrosis and steatosis, little brown granules in cytoplasm were showed in group H.2 Liver laboratory findingsThe value of ALT in groups A,B,C,D,E,F,G and H were 45.75±8.08U/L,432.12±116.01U/L,71.25±14.50U/L,105.75±32.66U/L,87.67±36.49 U/L,44.80±8.77U/L,50.00±8.73U/L and 48.20±8.99U/L; The value of AST in groups A,B,C,D,E,F,G and H were 144.50±22.56U/L,130.50±9.25U/L,173.00±21.71U/L,187.38±40.11U/L,149.00±30.78U/L,139.40±8.50U/L,135.70±11.22 U/L,728.63±335.36U/L. The value of TBIL in groups A,B,C,D,E,F,G and H were 0.06±0.19umol/L,25.28±21.18umol/L,3.15±3.79umol/L,2.59±3.37umol/L,3.23±3.3 umol/L,0.01±0.03umol/L,0umol/L,0.01±0.03umol/L; The value of GGT in groups A,B,C,D,E,F,G and H were 4.47±3.12u/L,16.98±9.97u/L,3.79±2.28u/L,3.78±2.03 u/L,3.75±1.67u/L,2.05±1.66u/L,2.44±1.85u/L,1.17±1.13u/L. As for the levels of ALT,AST,TBIL and GGT in serum in Group B were significantly higher than that in groups A, C, D and E (p<0.05); No statistical difference exist among groups A, C, D, E, F, G and H (p>0.05).3 The measurement of HMGB1 in cytoplasmThe OD value of HMGB1 in cytoplasm were 10003.08±846.71,29711.96±7826.83,16092.66±2308.52,18948.78±4495.19,15221.1±4633.58,11348.65±1192.3,12172.38±1451.77 and 10829.43±1310.7 in groups A, B, C, D, E, F, G and H. As for the levels of HMGB1 in model groups were significantly higher than that in the drug prevention groups and treatment groups (p<0.05); No statistical difference exist among other groups (p>0.05).4 The correlation between the liver tests and HMGB1The coefficient between ALT and HMGB1 was 0.922 and the coefficient between AST and HMGB1 was 0.829, which meant linear correlation (p<0.05); There were no correlation between TBIL, GGT and HMGB1.Conclusion:1 HMGB1 played a role in DILI inflammatory process as a inflammatory factor.2 HMGB1 and the liver function index showed a proportional relationship.3 S-adenosylmethionine and diammonium glycyrrhizinate can restore the DILI damage by inhibiting HMGB1 expression and the effect of drugs combination was not superior to separate use for the expression of HMGB1.
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