The Study About The Changes Of Hepatic Pathology And Hepatic Fibrosis In The Children With Biliary Atresia After Kasai Portoenterstomy

Objective 1. To investigate the hepatic characteristically of pathological changes in the children with biliary atresia(BA) after Kasai portoenterostomy(KP), and analysis the relation between the degree of hepatic damage and cholangioitis. 2. To investigate surface glycoprotein protein of hepatocyte 14(CD14), vascular endothelial growth factor 34(CD34), transforming growth factor-β1(TGF-β1) expression in liver biopsy of biliary atresia, the aim of this study is to explore its role of liver fibrosis process in biliary atresia.Methods 1. A retrospective study was conducted in32 consecutive patients with BA between May 2012 and December 2013, 16(10male and 6 female) patients of this group were received liver transplantation and removal native liver from Tianjin First Center Hospital. The other 16 patients(11male and 5 female) of this group received KP, then taken specimens from the liver during operation in Tianjin Children’s Hospital. The pathology of the liver tissue from these two groups was studied. Pathological study included Hematoxylin and eosin(HE) staining and Immunohistochemical(IHC) staining for evaluating the grade of liver fibrosis, extension of bile duct and blood vessels proliferation, degree of hepatocellular damage and infiltration of inflammatory cells.2. From January 2013 to January 2014, we collected liver biopsy from BA patients who received Kasai procedrue(Kasai group) in Tianjin Children’s Hospital and the BA patients after Kasai procedure who received liver transplantation(transplantation group) in Tianjin First Center Hospital, the liver biopsy from congenital biliary dilation patients(the CBD group, is set to control) during the surgery in in Tianjin Children’s Hospital, 15 cases in each group. Use HE staining to observe the liver fibrosis degree; CD14, CD34, TGF-β1 immunohistochemistry were used to detect protein expression and localization in liver tissue, and analyze the positive expression relationship between the three proteins in the liver biopsy. Then we compared the role in the progress of liver fibrosis in different groups. Application of computer imaging systems to collect photos, image analysis software(Image-Pro-Plus) were used to analysis CD14, TGF-β1 protein integrated optical density(IOD) in different groups,and CD34 positive area(Area). All the data using SPSS17.0 statistical software for single factor ANOVA analysis(p < 0.05), and want to know whether protein expressed in three groups has any difference statistically, Pearson Correlation analysis(p < 0.01) showed us if there is correlation between the three groups about the expression of different protein.Results 1. The appearance of the liver in the control group was swelling, severe cholestasis and medium texture. The Grading of liver fibrosis was between II ~ IV, the portal bile plugs was 0 ~ I, the proliferation of blood vessels is III ~ IV, bile duct’s proliferation is I ~ III. Macroscopically, the liver of experimental group had more atrophy areas in the liver. Grading of liver fibrosis was divided to level IV, portal bile plugs was I- II, the proliferation of portal blood vessel IV, the proliferation of bile duct proliferation was divided to III ~ IV. 2. According to the experimental results, CD14 was expressed in myeloid liver sinusoidal endothelial cells, fibroblasts and the inflammatory cells. TGF-β1 expressed in fibroblasts, biliary epithelial cells, inflammatory cells. CD34 expressed in the portal area and the proliferation of vascular endothelial cells. Compared with the CBD group, the expression of CD34, CD14, TGF-β1 proteins express high expression in the experimental group. From the perspective of liver fibrosis grading(Ⅰto Ⅳstage) in biliary atresia, the expression of CD14, TGF-β1 protein decreased gradually, the expression of CD34 increased. The difference between the three groups about the expression of CD14, TGF-β1, CD34 proteins was statistically significant(P<0.05), the expression of CD14, TGF-β1 proteins were significantly correlated(P<0.05), whereas the expression of TGF-β1, CD34 proteins was significantly positive correlated.Conclusion 1. The liver damage of biliary atresia was impacted by the time of Kasai portoenterostomy(KP), the patients who recived early KP, hepatic vascular and bile duct were hyperplasia obvious, as well as a slight lobular cholestasis; late KP, the liver were cholestatic seriously in patients, pseudolobule bridge fibrosis, bile ducts were hyperplasia obvious. After KP, biliary atresia patients who underwent liver transplantation were at end-stage liver fibrosis and severe cholestasis, bile ducts and vasculars proliferation severe, periportal widened, inflammatory cells infiltrate.2. The expression of CD14, TGF- beta 1 in liver fibrosis progress are increased gradually in BA, are involved in BA liver fibrosis, and they all can accelerate the liver cirrhosis of BA. TGF-β1 and CD34 were significantly positively related in liver tissue in three groups, they may interacted with each other in fibrosis, and played a role in the progression of fibrosis. wheras in advanced disease,all the cells are damaged and were substituted by fibrosis, three proteins are gradually decline.