The Study On The Sensitivity Of HT1Model Mice To Alcoholic Liver Damage

Alcoholic liver damage seriously influences the human health, and the mechanism of alcoholic liver damage needs further research, whether hereditary tyrosinaemia type I model mouse is sensitivity to alcohol has not been reported.ObjectiveUsing Fah knockout mice to establish alcoholic liver damage model, and to detect the mechanism of alcoholic liver damage, to provide the basis for preventing and controlling the alcoholic liver damage.Materials and methodsTwenty male Fah knockout (Fah－/-) and twenty male Fah wild type (Fah+/+) mice were got through the genotype identification of the descendants which were generated by129mice mating. These two genetic types of mice were randomly divided into two control groups and two model groups, ten mice per group. According to Liber-Decarli model to set up alcoholic liver damage model, while the mice of two control groups were given equal liquid diet without alcohol.After feeding3weeks, the levels of serum ALT, AST and TG were detected. Then the mice were condemned to death, the livers were dissected and weighed to calculate the liver index. Part of the liver tissue was prepared to make the10%liver tissue homogenate for the MDA, SOD and GSH levels detection. Part of the liver tissue was fixed with paraffin to do HE and TUNEL staining. The extent of liver damage and the liver cell apoptosis were examined. The mRNA expression levels of IL-6,TNF-a,TGF-β,Caspase-8,Bax,Bcl-2were detected by Real-time PCR technology. The protein expression levels of caspase-8,Bax,Bcl-2were detected by Western-blot technology.The Repeated Measurement ANOVA and One-Way ANOVA were carried out to analyze the difference by use of the SPSS11.5software, measurement date were expressed with x-S. The significant level was set at α=0.05.Results1The changes of body weight and liver index:The mice weight of the two control groups increased, while those of the two model groups decreased(P<0.05). Compared with their control groups, the liver index of the two model groups increased(P<0.05); The liver index of the Fah-/-mice model group were higher than those of the Fah+/+mice model group(P<0.05).2The changes of mice serum biochemical index and liver fat:The ALT,AST and TG levels of the two control groups were not statistically significant (P>0.05); The ALT, AST and TG levels of the two model groups were higher than those of their control groups(P<0.05); The ALT, AST and TG levels of the Fah-/-model group were higher than those of the Fah+/+model group(P<0.05).3The changes of mice liver oxidation and antioxidation levels:The MDA,GSH and SOD levels of the two control groups were not statistically significant (P>0.05); The MDA levels of the two model groups were higher than those of their control groups(P<0.05), the GSH and SOD levels were lower than those of their control groups(P<0.0.5); The MDA levels of the Fah-/-model group were higher than those of the Fah+/+model group(P<0.05), the SOD levels were lower than those of the Fah+/+model group(P<0.05); The GSH levels of the two model groups were not statistically significant (P>0.05).4The changes of liver pathology: Compared with the control groups, the mice liver size increased, lustre grayed,normal liver tissue structure disappeared, liver cell cable arranged disorderly, cytoplasm loosed, fat empty bubble and the balloon degeneration can be seen in cytoplasmic in the two model groups. The liver damage of the Fah-/-model group were more serious than those of the Fah+/+model group.5The mRNA and protein expression levels of related gene:The TNF-α,IL-6,TGF-β, Caspase-8,Bax and Bcl-2mRNA levels between the two control groups were not statistically significant (P>0.05), The TNF-α and IL-6mRNA levels of the two model groups were higher than those of their control groups(P<0.05), the Bcl-2mRNA levels were lower than those of their control groups(P<0.05); The TNF-a,IL-6and Caspase-8mRNA levels of the Fah-/-mice model group were higher than those of the Fah+/+mice model group(P<0.05), the Bcl-2mRNA levels were lower than those of the Fah+/+mice model group(P<0.05), the TGF-β and Bax mRNA levels between the two model groups were not statistically significant(P>0.05). Western-blot results showed that the Caspase-8and Bax protein levels of the two model groups increased and Bcl-2decreased obviously; The Caspase-8and Bax protein levels of the Fah-/-model group were higher than those of the Fah+/+model group(P<0.05), the Bcl-2protein levels were lower than those of the Fah+/+model group(P<0.05).6The cell apoptosis:Tunel staining showed that the liver cell apoptosis of the Fah-/-mice model group was more serious than that of the Fah+/+model group.ConclusionsThe chronic alcoholic liver damage model was successfully established by sustained feeding mice with alcoholic liquid; The knockout of Fah may aggravate alcoholic liver damage; Alcoholic liver damage is related to oxidative stress, inflammatory reaction and liver cell apoptosis.