| Objective Establish the mouse model of acute alcoholic liver injury to investigate the effects of obeticholic acid on acute alcoholic liver injury.Methods Established the model of acute alcoholic liver injury in mice.Mice were randomly divided into 8 groups(6 per group): Control-2 h,8 h group,OCA-2 h,8 h group,Alcohol-2 h,8 h group,OCA + Alcohol-2 h,8 h group.From the first day to the fifth day,OCA-2 h and 8 h groups and OCA + Alcohol-2 h,8 h groups mice were given OCA(10 mg/kg)once a day by gavage,five times in total;Control-2 h,8 h groups were given the same volume of saline;from the third day,alcohol-2 h,8 h groups and OCA + alcohol-2 h,8 h groups were given 50% alcohol(4 g / kg)twice a day by gavage,with an interval of 12 h;on the fifth day,mice were given alcohol once,five times in total.After the last administration of alcohol by gavage,the mice were killed painlessly after fastting for 2 hours or 8 hours respectively,and the weight of mice and liver were weighed,the serum was taken to detect the related biochemical indexes.Some liver tissues were stained with H&E to observe the liver pathology,the other liver tissues were detected with RT-PCR to detect the transcription of TNF-?and IL-1?,and with Western blotting to detect the expression of HO-1 and CYP2E1.Results(1)The effect of pretreatment with OCA on liver coefficient of mice:after alcohol treatment,the liver coefficient of mice increased significantly in alcohol-2h,8 h group;but markedly decreased in OCA + alcohol-2 h,8 h group.(2)The effects of pretreatment with OCA on the gross morphology of mice liver: the liver of mice showed paleness,hyperemia and edema,but were substantially reduced in OCA + alcohol-2 h,8 h groups.(3)The effects of pretreatment with OCA on the serum ALT of mice: in alcohol group,the serum ALT of mice were increased in Alcohol-2h,8h groups,but decreased significantly after pretreating by OCA.(4)The effect of pretreatment with OCA on oxidative stress in liver of mice:in alcohol-2h,8h groups,the content of MDA increased,GSH and GSH-Rd decreased,but GST did not change significantly;after pretreating with OCA,the content of MDA decreased,the content of GSH and GSH-Rd increased,but the activity of GST did not change apparently.(5)The effect of pretreatment with OCA on liver pathology in mice:in the alcohol-2 h,8 h groups,the hepatocyte cords were disordered,the boundary were fuzzy,the shape and size were different,the hepatocytes were edematous,the local hepatocytes were swollen,ballooned,the single or scattered hepatocytes were necrotic,the liver tissues were congestive and inflammatory cell infiltration,but the degree of swelling,denaturation and necrosis of hepatocytes were significantly reduced in the OCA +Alcoho-2 h,8 h groups.(6)The effect of pretreatment with OCA on the pro-inflammatory gene of alcoholic liver injury in mice:m RNA level of hepatic TNF-? and Il-1 ? in alcohol-2 h and 8 h groups were increased and lower in OCA +alcohol-2h group,but there was no statistical significance.(7)The effect of pretreatment with OCA on the level of Cyp2e1 in the cytoplasm: the expression of Cyp2e1 were increased significantly in the alcohol-2 h,8 h groups,while decreased dramatically after OCA pretreatment in OCA + alcohol-2 h,8 h groups.(8)The effect of pretreatment with OCA on the level of HO-1 in the cytoplasm: the expression of HO-1 was increased significantly in the alcohol-2 h group,while decreased after OCA pretreatment in OCA + alcohol-2 h group.The expression of HO-1 was increased in the alcohol-8 h group,while decreased markedly after OCA pretreatment in OCA +alcohol-8 h group.Conclusion Obeicholic acid has a significant protective effect on acute alcoholic liver injury in mice.It may be related to the inhibition of the expression of Cyp2e1 and HO-1,the promotion of the recovery of antioxidant enzyme activity and the inhibition of lipid peroxidation. |
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