Establishment Of Acute Hepatic Injury Model In Mice And Analysis Of GSTA1

Recently, liver disease serious threaten to human healthy and the incidence of liver desease in aquaculture are increased significantly. Liver disease not only causes a serious economic damage, but also causes directly or indirectly harm to human through the food chain, which bring series of food safety issues. Acute hepatic injury that cause the development of disease and developing into liver failure finally is the initiating factor and common pathway of numerous liver disease. It is extremely important to establish animal model which disease process is similar with human acute hepatic injury. In depth research and investigate pathogenesis and treatment measures of acute hepatic injury is important to prevention of chronic liver disease. GSTA1is a subtypes of Ⅱ combination reaction isoenzyme GST, which has important protective effect on oxidative damage and tumor, and it is likely to be an important new direction for drug development. Currently, there are less investigation about change of GSTA1in acute hepatic injury and no research on the role of GSTA1when hepatic injury. To determine the change of GSTA1in acute hepatic injury is the basis to investigate protective effect and mechanism of GSTA1.The research will establish three mice model of acute liver injury based on the optimum exposure pathway, dose and time selected by research. Evaluate the CCl4-induced model by levels of serum transaminase(ALT, AST), liver homogenates indicators (SOD, MDA, GSH, GSH-Px) and liver histology analysis. Evaluate the APAP-induced model by the levels of serum transaminase, liver homogenates indicators (SOD, MDA, GSH, GSH-Px, NO) and liver histology analysis. Evaluate the ethanol-induced model by the levels of serum transaminase, liver homogenates indicators (SOD, MDA, GSH, GSH-Px, TG) and liver histology analysis. The change of concentration and activity of GSTAl in serum and liver were detected in model replicated successfully. The concentration of GSTA1was detected by mouse GSTA1kit which is Elisa principle. The activity of GSTA1was detected by NBD-C1which is the substrate of GSTA1catalytic reaction and calculated by the reaction product NBD-SG base on colorimetry. Enzyme activity is expressed as the amount of reaction products of GSH of per minute per milliliter in serum. Enzyme activity is expressed as the amount of reaction products of GSH of per minute per mg of cytosolic protein in liver.The result shows:1. The mice model of acute chemical hepatic injury can be set up by gavage0.35%carbon tetrachloride,10mL·kg-1, for24h. The change of serum transaminase and all liver indicators in model group has significant differences compare with control group (P<0.01). Liver histopathologic study shows hepatocyte granular degeneration, cytoplasm disintegration, nuclear condensation and inflammatory cell infiltration in model group. The concentration and activity of GSTA1has significantly increase(P<0.O1)in serum, however, it has significantly reduce (P<0.01) in liver.2. The mice model of acute drug-induced liver injury in mice can be set up by gavage200mg·kg-1body weight APAP, for12h. The change of serum transaminase and all liver indicators in model group has significant differences compare with control group (P<0.01). Liver histopathologic study shows congestion congestion, inflammatory cell infiltration, hepatocyte cord fuzzy, nuclear condensation in model group. The concentration and activity of GSTAl has significantly increase (P<0.01) in serum, however, it has significantly reduce (P<0.01) in liver.3. The mice model of acute alcoholic liver injury in mice can be set up by gavage50%ethanol,14mL·kg-1, for8h. The change of serum transaminase and all liver indicators in model group has significant differences compare with control group (P<0.01).Liver histopathologic study shows round vacuoles, inflammatory cell infiltration, hepatocyte spotty necrosis in model group. The concentration and activity of GSTA1has significantly increase (P<0.01) in serum, however, it has significantly reduce (P<0.01) in liver.My research was successfully replicate the animal model of acute chemical, drug, and alcoholic hepatic injury. The concentration and activity of GSTAl has significantly increase (P<0.01) in serum, and has significantly reduce (P<0.01) in liver. The results indicated that GSTA1in liver can released into the blood to participate in the antioxidant activity and protect liver when acute hepatic injury. The change of GSTA1in serum indicated that GSTA1has important significance to evaluate the model. This research may lay a foundation for further investigation on regulatory role of hepatoprotective drug to GSTA1and the molecular mechanisms of GSTA1.