The Expression And Significance Of The TGF-β1 Signaling Pathways Related Protein In Biliary Atresia

Objective:To Study the expression of transforming growth factor beta 1(TGF)-beta 1 related regulatory proteins and direct product,integrin alpha v 8 、 p38 、 the extracellular regulatory protein kinase 2(ERK2)、c-Jun N-terminal kinase(JNK2),tissue inhibitor of metalloproteinase 1(TIMP-1)and Collagen Ⅲ protein in the liver of children with biliary atresia(BA),and to explore the alpha v beta 8,p38,ERK2,JNK2 and TIMP-1 and Collagen Ⅲ the function in the process of liver fibrosis of biliary atresia.method:Liver biopsy specimens were collected from Tianjin Children’s Hospital from January 2013 to October 2016 who undertook Kasai procedure because of biliary atresia named BA group(n=20)and who had liver biopsy duo to congenital biliary dilatation(CBD,n=20);he transplantation group(n=20)were collected from Tianjin first central hospital(who need liver transplantation after Kasai procedure due to liver failure and other related causes of liver transplantation.The liver tissue samples were taken from the front of the right hepatic lobe,and were performed by hematoxylin and eosin(HE)stain、the immunohistochemical(IHC)staining and quantitative real-time polymerase chain reaction(qRT-PCR),HE stain were used to observe liver cell changes and inflammatory cell distribution,IHC were used to detect the expression of αvβ8,p38,ERK2,JNK2,TIMP-1 and CollagenⅢ in liver tissue of biliary atresia,qRT-PCRwas used to test the gene expression of αvβ8,p38,ERK2,JNK2,TIMP-1 and CollagenⅢ.The application of image analysis software(Image-Pro – Plus,IPP)and SPSS 15.0 software Image to analysis differences of αvβ8,p38,ERK2,JNK2,TIMP-1 and CollagenⅢ in the group of CBD,BA and liver transplantation group,and to explore the function in the process of liver fibrosis of biliary atresia.Results:1.HE staining.in the CBD group,there are a little fiber cell hyperplasiaa and a small amount of inflammatory cells infiltration;in the BA group,portal area was widened,a large number of inflammatory cells infiltration,the fibrous tissue hyperplasia was obvious,and the phenomenon of bridging fibrosis was common,and the pseudolobule formation was observed;in the liver transplantation group,the portal area was obviously broadening,fibrous tissue hyperplasia serious,extensive bridging fibrous tissue formation,the pseudolobule were obviously,scar tissue formation were distinctly.2.immunohistochemistry.(1)The qualitative analysis of immunohistochemical:the main positive parts of these proteins are liver cell cytoplasm,portal area bile duct epithelial cells and vascular endothelial cell cytoplasm.1)αvβ8 protein.The αvβ8 protein was stained in the cytoplasm of liver cells,portal area bile duct epithelial cells and vascular endothelial cells cytoplasm,a weakly expressed in the CBD group,and in the BA group and liver transplantation of liver cell cytoplasm,collect abbacy bile duct epithelial cells and endothelial cell cytoplasm was strongly positive expression.2)p38 protein.The p38 protein was stained in the cytoplasm of liver cells,portal area bile duct epithelial cells and vascular endothelial cells cytoplasm.Above cells were negative expression in the CBD group,however the positive cells were strong positive expressed in the Kasai group and the liver transplantation group.3)ERK2 protein.In the CBD group,weakly positive expression in hepatic sinus surrounding cells and endothelial cell cytoplasm,in BA group and the liver transplantation group,not only strongly positive expression in the vascular endothelial cells and hepatic sinus surrounding cells,but also strongly positive expression in the bile duct epithelial cells and hepatocytes cytoplasm.4)JNK2 protein.In the three groups of liver tissue,JNK2 protein was positive expression in hepatocyte cytoplasm,portal area bile duct epithelial cells and vascular endothelial cell cytoplasm,in the CBD group they were weakly positive expression,however in the BA group and liver transplantation group they were positive expression.5)TIMP-1 protein.The positive expression cells of this protein in the liver tissue were the cytoplasm of the hepatocytes,the cytoplasm of the vascular endothelial cells and the cytoplasm of the bile duct epithelium cells in the portal area.In the CBD group these cells were weakly positive expression,but in the BA and liver transplantation groups,liver tissue samples they were positive expression.6)Collagen III protein.In the group of CBD,in the liver tissue,the Collagen III protein was weakly positive expression in the cytoplasm of vascular endothelial cells,bile duct epithelial cells and hepatocytes,while in the BA group and liver transplantation group these cells were Positive expression.(2)Immunohistochemical semi-quantitative analysis:in three groups,the expression of αvβ8,p38,ERK2,JNK2,TIMP-1 and CollagenⅢ in BA group and liver transplantation group were significantly higher than those in the CBD group(p <0.05).However the content of αvβ8,p38,ERK2,JNK2,TIMP-1 and CollagenⅢ had no significant difference between the transplanted group and the BA group(p> 0.05).3.qRT-PCR.The mRNA content of αvβ8,p38,ERK2,JNK2,TIMP-1 and CollagenⅢ in BA and transplantation group was significantly higher than those in group CBD(P <0.017).However,there was no significant statistical difference in BA and liver transplantation(P> 0.05).Conclusion:In the children with biliary atresia,along with the development of the disease and the increased degree of the liver fibrosis,the expression of αvβ8,p38,ERK2,JNK2,TIMP-1 and CollagenⅢ protein in BA liver tissue was significantly increased,which indicated that αvβ8,p38,ERK2,JNK2,TIMP-1 and CollagenⅢ proteins may be involved in and promote BA hepatic fibrosis process.