| Objective:From the experiment,Establish a model of hepatic ischemia reperfusion injury(HIRI)after aotologous orthotopic liver transplantation(AOLT)in rats,to determine whether pretreatment with Xuebijing can improve HIRI after liver transplantation,and explore the specific mechanism of action of Xuebijing in HIRI after liver transplantation.Methods:(1)60 SD♂rats were randomly divided into blank group(A),HIRI group(B),and HIRI pretreatment group(C)to establish HIRI model after AOLT operation in rats;Before operation,groups A and B were pretreated with normal saline and group C with Xuebijing respectively.Rats in experiment A only opened the intestine after laparotomy,without liver transplantation,and groups A and B received AOLT.(2)Record the rat liver-free period and operation time,and postoperative survival rate.The rats were sacrificed 1d,2d,3d,and 7d after the experiment,and serum and liver tissue samples were obtained.(3)Liver function tests of ALT,AST and TBIL are carried out by automatic biochemical analyzer.(4)Detection of tumor necrosis factorα(TNF-α)and interleukin(Interleukin-6,IL-6)in serum by ELISA.(5)Liver tissues were collected for hematoxylin-eosin(HE)staining,observing the pathological changes of liver tissue under light microscope.(6)Western blotting was used to detect the expression of Bax and Bcl-xl proteins in liver tissue.(7)Immunohistochemical detection:Observe the expression of Bax and Bcl-xl protein in liver tissue under microscope.(8)The obtained data is expressed as mean±standard deviation(X~-±S),using SPSS20.0 statistical software to analyze the data,and carry out homogeneity test of variance,P<0.05 has statistical significance.Results:(1)Comparing the two groups of B and C,there was no significant difference in liverless period and operation time(P>0.05).(2)On days 1,2,3,and 7after surgery,the serum ALT,AST,and TBIL indexes in groups B and C were significantly higher than those in group A,the numerical value of group C is lower than that of group B between the two groups,compared with each other(P<0.05).(3)Analysis of inflammatory factors in the serum of three groups of rats showed that Group A had the lowest levels of TNF-αand IL-6,the levels of B and C increased,the level of Xuebijing group C was significantly lower than that of group B.(4)Observing the pathology of liver tissue under light microscope,the liver tissue structure and morphology of group A were normal without obvious congestion,group B liver blood sinus congestion,spotted,focal necrosis area,inflammatory cell infiltration can be seen.The liver tissue lesions in group C were significantly alleviated compared with the control group.(5)Western Blot analysis:Using Image J software to analyze the band gray scale,Bax protein is the lowest expressed in A,and is highly expressed in B and C,but C is less than B.(6)The Bax/Bcl-xl ratio in group C was lower than that in group B(P<0.05).Conclusion:1、Xuebijing can reduce the levels of ALT,AST,TBIL,TNF-α,IL-6 in serum to protect liver function after hepatic ischemia reperfusion injury.2、Xuebijing can regulate the expression of Bax and Bcl-xl proteins,prevent hepatocyte apoptosis,and thus protect liver tissues. |
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