The Effects Of Tunicamycin Preconditioning On Isoniazid-induced Liver Injury In Rats And Its AMPK Phosphorylation

Objective(1)To establish isoniazid-induced liver injury model in rats,explore the possible mechanism of isoniazid-induced hepatic steatosis by studying the changes of AMPK phosphorylation level,and its downstream target gene expression in this model.(2)investigate the role of tunicamycin Pretreatment on isoniazid-induced liver injury in rats and its AMPK phosphorylation.Methods(1)24 SPF male SD rats were randomly divided into three groups,control group and low-dose model group(50 mg/kg)and high-dose model group(100 mg/kg).Each group consisted 8 animals.Gastric treatment with Isoniazid.The rats were sacrificed after 2 weeks.The liver index and serum biochemical indexes of ALT?AST?TBIL?DBIL?TBA?TG?TC and liver TG content were measured.The histopathological changes of liver was observed.The expression of p-ACC was detected by immunohistochemistry.AMPK?p-AMPK and FAS protein expression were detected by Western blot.(2)32 SPF male SD rats were randomly divided into Four groups,normal control group,model group,experimental group and experimental control group.Each group consisted 8 animals.Isoniazid 100 mg /(kg · d)was given to the rats in the experimental group and the experimental group,tunicamycin was intraperitoneal injection 200?g /kg pretreated.The rats were sacrificed after 2 weeks.the liver index and serum biochemical indexes of ALT?AST?TBIL?DBIL?TBA?TG?TC and liver TG ?TC content were measured.IRE1??p-IRE1??XBP1? AMPK?p-AMPK protein expression were detected by Western blot.Results(1)Compared with normal control group,liver index,serum ALT,AST,TG and liver TG content in the high-dose model group were significantly increased(P <0.01 or P<0.05);Pathological changes manifested as steatosis,necrosis,inflammation.The expression of p-AMPK and p-ACC was reduced,the expression of FAS was increased.(2)Compared with normal control group,liver index,serum AST,TBA and TC in the model group increased significantly(P<0.01).Compared with model group,liver index,serum ALT,AST,TC,TG in the treatment group were significantly increased(P <0.01 or P<0.05).the liver TG content in the treatment group increased,the liver TC content in the treatment group increased(P<0.05).Pathological changes manifested as steatosis,necrosis,inflammation in the model group.The treatment group Pathological changes more seriouily.The expression of p-IRE1?/IRE1? in the treatment control group was significantly increased than the normal group(P<0.01).Compared with normal group,The expression of p-IRE1? ?XBP1 and p-IRE1?/IRE1? ratio in the model group was increased,the expression of p-AMPK and the p-AMPK/AMPK ratio was decreased(P<0.01).Compared with the model group,the expression of p-IRE1??XBP1 and p-IRE1?/IRE1? ratio in the treatment group was increased,the expression of p-AMPK and p-AMPK/AMPK ratio was decreased(P<0.01).Conclusion(1)The isoniazid-induced liver injury may be related to the inhibition of AMPK,ACC phosphorylation and subsequently promotion of FAS.(2)Isoniazid induced endoplasmic reticulum stress activation during liver injury in rats(3)Tunicamycin Pre-treatment aggravate isoniazid-induced liver injury in rats,which could be related to the inhibition of AMPK via Endoplasmic Reticulum Stress.