Experimental Study On The Role Of Bile Duct Epithelial Mucin In Ischemic Type Biliary Lesion After HAE

Objective:Establishment of a new animal model of ischemic tube injury after HAE,To investigate the expression and relationship of mucin in bile duct epithelial cells in ischemic bile duct injury after HAE.Methods:Part one:New Zealand white rabbits were randomly divided into group A(n=15),group B(n=20)and group C(n=20).ALP was detected by ELISA method,and the levels of y-GT and TBil were detected by colorimetry to observe the degree of bile duct injury.The lipiodol deposition was observed by scanning the lipiodol liver of experimental rabbits with CT instrument.The histopathological changes of bile duct were observed by hematoxylin-eosin(HE)staining.Part two:New Zealand white rabbits were randomly divided into control group(sham group)and operation group(surgery group)with 5 rabbits in each group.Samples were taken 24 hours after operation.The gene expression of Muc1,Muc2,Muc3A,Muc4 and Muc5AC was detected by Real time-PCR method,and the protein expression of Muc1 and Muc5AC was detected by Western Blotting method.Result:1.Establishment and evaluation of ITBL animal model after HAE:Under the method of modified microsurgical intubation and hepatic artery reconstruction,an animal model of ischemic bile duct injury after HAE was established.The success rate of operation in group C was 85%(17/20),and the postoperative survival rate was 88.2%(15/17).The serum levels of ALP,y-GT and TBil in group C at 1 day,3 days and 7 days after operation were significantly higher than those in group An and group B(P<0.05).The results of HE staining showed that the bile duct epithelium in group C showed different degrees of disorder of cell polarity,cell exfoliation,cytoplasmic porosity,vacuolation,punctate necrosis and a small amount of focal necrosis on the 1st,3rd and 7th day after operation.2.Changes of mucin expression in bile duct epithelial cells in ITBL after HAE:The results of Western blotting showed that the expression of Muc5AC protein in bile duct epithelial cells in the operation group was significantly lower than that in the Sham group,this is consistent with the change of gene level.The results of Real time-PCR experiment showed that compared with Sham group,the expression of Muc3A mRNA decreased significantly,but there was no significant change in Muc2 mRNA and Muc4 mRNA.Conclusion:1.Using the modified microsurgical method of hepatic artery intubation and hepatic artery reconstruction,we can successfully establish an animal model in accordance with the pathophysiology of ITBL after clinical TACE or HAE.It is an ideal animal model for the study of biliary complications after hepatic intervention.2.The decreased expression of mucin Muc3A mRNA and Muc5AC in bile duct epithelial cells after HAE may be related to ischemic bile duct injury after HAE,while the expression levels of mucin Muc1,Muc2 mRNA and Muc4 mRNA in bile duct epithelial cells are limited by HAE,which may not be related to ischemic bile duct injury after HAE.