| Objective: To investigate the role of the Inflammatory Cytokines in drug-induced liver injury (DILI) process in DILI model rat generated with carbon tetrachloride administering and evaluating the role of diammonium glycyrrhizinate (DGL) and S-adenosylmethionine (SAMe) in the treatment of Drug-Induced Liver Injury.Methods: 25 male and 25 female wistar rats weighting 200±20grams were purchased from the expremental animal center of Hebei medical university. Rats were acclimatized for 7 days and randomly divided into groups A, B, C, D and E with 10 animals each group. Group A was given normal saline alone by intragastric administration. Groups B, C, D and E were separately given 5ml/kg of 50% carbon tetrachloride at first, and 2.5ml/kg of 50% carbon tetrachloride by subcutaneous injection subqently with 2 times per week for four weeks to generate drug-induced liver injury animal model. Group C was given diammonium glycyrrhizinate 375mg/kg/d, Group D was given S-adenosylmethionine 0.833g/kg/d and Group E was given diammonium glycyrrhizinate 375mg/kg/d plus S-adenosylmethionine 0.833g/kg/d by intragastric administrati- on simultaneously. Two rats died in 4th week in Group E. Two rats in Groups A, B, C, D and E were fasted and sacrificed randomly at the end of the second week, the others were fasted and sacrificed at the end of the fourth week, and the serum were collected. The liver function marker of alanine aminotransferase (ALT), aspartate amino transferase (AST), albumin (ALB) and total bilirubin (TBIL) in serum were measured with automatic biochemistry analyzer. Pathological examination was performed with hematoxylin eosin (HE) section. The levels of tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) in serum were measured with radiation immunity arithmometer. The levels of adiponectin in serum were measured by ELISA.Results:1 The status of Wistar rats The mental status of group B is weak and the fur is dry.The level of body weight in group B was lower significantly than that of other groups(p<0.05), The wistar rats in group B is characterized with slow movements, drowsiness, slow growth and behavioral disorders such as rolled and restless moving. The mental status of Group A is good, and the fur is thick and smooth. They have comfortable posture and normal body weight.the status of other Groups are better than group B but worse than group A.2 Liver histopathological examination: the examination of hepatic pathology in groups A, B, C, D and E at the 2th week: the section of group A showed normal liver cells with normal hepatic plate and no inflammatory cell infiltrating; Obvious swelling of liver cells, crowding of hepatic plate, scattered steatosis and piecemeal necrosis, sinus hepaticus with mild hematoma expansion were showed in Group B; Mild swelling of liver cells, scattered steatosis and piecemeal necrosis were showed in Group C; swelling of liver cells, diffuse distribution of steatosis and piecemeal necrosis, crowding of hepatic plate in Group D; while mild swelling of liver cells, diffuse distribution of steatosis, scattered piecemeal necrosis in Group E.The examination of hepatic pathology in all groups at the 4th week: the section of group A showed normal liver cells with normal hepatic plate and no inflammatory cell infiltrating; Obvious swelling of liver cells, diffuse distribution of piecemeal necrosis, obvious capillary bile thrombus and activing mesenchymal reaction showed in Group B; Swelling of liver cells, obvious piecemeal necrosis and scattered steatosis in Group C; Mild swelling of liver cells, scattered steatosis and piecemeal necrosis, partial liver cytoplasm pigment deposition in Group D; Obvious swelling of liver cells, scattered steatosis, piecemeal necrosis and partial liver cytoplasm pigment deposition in Group E.3 The measurement of ALT, AST, ALB and TBIL in serum We studied the liver function in DILI model animal, compared with groups A, C, D and E, the level of ALT, AST and TBIL in serum in Group B was significantly higher (p<0.05), respectively. Compared with groups A, C, D and E, the level of ALB in serum in Group B was lower significantly (p<0.05); no statistical difference exist refered to ALT, AST, ALB and TBIL levels among groups A, C, D and E (p>0.05).4 The measurement of TNF-α, IL-6 and Adiponectin in serumThe contents of TNF-αin serum is 1.27±0.18, 1.65±0.18, 1.26±0.18, 1.28±0.26 and 1.26±0.23ng/ml in groups A, B, C, D and E respectively. Compared with groups A, C, D and E, the level of TNF-αin Group B was significantly higher (p<0.05); no statistical difference exist for TNF-αlevels in groups A, C, D and E (p>0.05).the contents of IL-6 in serum is 38.01±9.36, 43.19±26.16, 33.41±8.62, 40.93±20.37 and 28.39±11.01pg/ml in groups A, B, C, D and E respectively. No statistical difference exist for IL-6 levels in groups A, B, C, D and E (p>0.05).The contents of Adiponectin in serum is 322.73±65.21, 406.73±193.84, 342.08±103.59, 429.66±247.56 and 296.89±34.72ng/ml in groups A, B, C, D and E respectively. No statistical difference exist for Adiponectin levels in groups A, B, C, D and E (p>0.05).Conclusion:1 The model of drug-induced liver injury can be successfully generated by subcutaneous injection of carbon tetrachloride. 2 TNF-αparticipate and have a role in DILI process, while IL and adiponectin do not invole or have not a important role in this process.3 S-adenosylmethionine and diammonium glycyrrhizinate can restore the DILI damage and liver function of DILI rat probably through inhibiting TNF-αexpression.
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