| Backgrounds and Objectives:Excessive drinking can cause multiple organ damage,of which the most significant is liver damage.Alcoholic liver disease is one of the leading causes of end-stage liver disease.Alcoholic liver disease has become the first leading cause of liver disease to death.As the alcohol consumption extended to the younger and older,it has made a profound influence for various age groups.Due to the differences of liver function among different age stages,we assume that there should have some differences between the degree of alcoholic caused liver injury.So we designed experiment models on different age stages of rats to study the effect of age factor on alcoholic liver injury.At present,the treatment of alcoholic liver disease is limited,studies also don’t have much progress.With the study of own stem cells,we proposed to study the feasibility of the stem cells in treatment of alcoholic liver injury and the possible mechanism by intravenous itself bone marrow mesenchymal stem cells.This experiment clear the following questions: 1.Age factors on acute alcoholic liver injury and liver cell apoptosis2.The effectiveness of BMSC in treating of alcoholic liver damage and related mechanism research.Methods:1.In this experiment,we feed 45 SD rats aged 1,6,18 months respectively,with 15 rats for each age.Then we randomly divided these ages into experimental group(n=10)and the control group(n=5).The experimental group members were treated with 15 g/kg40% ethanol by gavage,on the other hand,the control group were treated with the same dose of normal saline by gavage.After 48 h of gastric perfusion,collect the blood from the rat heart to measure the liver function.Samples of the liver tissue were taken to detect liver inflammation factor expression by ELISA,and to detect the rate of liver cell apoptosis by TUNEL separately.Then A further study on the expression of Bcl-2、Bax and Caspase3 in the apoptotic pathway after the alcohol stimulates in the liver was conducted to characterize the effect of age factor on acute alcoholic liver injury and its mechanism on the apoptosis of liver cell.2.In this experiment,we randomly divided 90 male SD rats weight 200-220 g intothe blank group,the control group and the BMSC group,with 30 in each group.The rat in the control group and BMSC group were gavaged with 15 g/kg 40% ethanol disposable lavage.After 12 hours,the BMSC group rats were injected 1ml normal saline which contains 5×10~6 BMSC cell by tail vein injection while the rats of control group were injected 1ml normal saline without any cells by tail vein injection.,samples of the liver tissue and the blood of the rats were taken to observe the liver function and liver inflammatory cytokines and the liver cell apoptosis respectively,1 day,3 days and 7 days after the rat tail vein injection.In order to research the effect of the BMSC treated the acute alcoholic liver injury in rats from the general situation of rats and the possible mechanism,we observed the rats survival rate,the liver function and inflammatory cytokines and the liver cell apoptosis.Results:1.In the control group,the liver function and inflammatory cytokines have no significant difference between the different age groups,the liver cell apoptosis rate between 6 month and 1 month group have no significant difference,while the rate of 18 month group is higher than the other two groups.In the experimental group,the liver function,inflammatory cytokines and the rates of liver cell apoptosis were significantly higher than the control group.And it is different among the different age groups: the expression of inflammatory cytokines in the 6 month group is highest.In the experimental group,the rate of apoptosis in 1 month is highest,the rate of 6 month is lower,while the rate of 18 month is lowest.The further study shows that the expression of Bcl-2,Bax,Caspase-3 is different in different age groups,and the result is in consistent of the trend of the liver cell apoptosis.2.Compared with the blank group,the rats activity and survival rate is reduced in control group and BMSC treatment group,and the liver function,inflammatory cytokines and the apoptosis rate are increased significantly(P<0.05).After 1 day and 3 days,compared with the control group,the liver function index,inflammation cytokines and apoptosis of the liver tissue were significantly decreased in the BMSC group,and the survival rate improved.After 7 days,the liver function,inflammatory cytokines and apoptosis of all groups decreased significantly compared with the result of 1day and 3days.Conclusions:1.In the physiological state,liver function and inflammatory cytokines have no significant correlation with age,and the rate of liver cell apoptosis increases with age.Acute excessive drinking can lead liver damage and liver cell apoptosis,and the damage degree and apoptosis rate are correlated with the age: the liver injury of the 6 month group is heavier,and the rate of apoptosis in 1 month group is higher.This may be related to the fact that different age rats under the stimulus of alcohol,the activation of liver inflammatory cell and hepatocyte apoptosis pathway is different.2.Alcoholic liver disease can cause significant increase of the liver function,inflammation cytokines,oxidative stress and cell apoptosis of rats;The resilience of rat liver is very strong,1 week after the liver injury by the alcoholic,liver function and inflammation factors recovered significantly.The injection of BMSC can improve the survival rate of the early alcoholic liver damage,and it can reduce the degree of liver tissue inflammation and cell apoptosis.Results of this research indicate that the BMSC may have the effect of treatment to alcohol on liver injuries.But the experiment shows that it may have no obvious effect on afterward recovery. |
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