The Study Of The Access Of MicroRNA145-Adducin3 And GBP-1 On The Mechanism In The Pathogenesis Of Proliferation Disorder Of Biliary Atresia

Objective:To determine the expression of MicroRNA145,Adducin3 and GBP-1 in liver tissue of infants with biliary atresia,to investigate the role of MicroRNA 145-Adducin3 pathway and GBP-1 in the pathogenesis of biliary atresia and its mechanism.Methods:The clinical and pathological data of 60 patients with biliary atresia(Study Group)were selected from January 2013 to January 2016 in Shenzhen Children's Hospital.The control group were selected for those younger than 6-month-old of non-cholestatic liver disease with normal liver function in 54 cases.The first part:Lyse the liver tissues of infants in BA group and control group and retract total RNA,using mRNA RT-PCR,MicroRNA RT-PCR and qRT-PCR analysis to identify the expression level of ADD3 mRNA as well as MicroRNA-145 in both groups.The second part:Construct the lentivurs with over-expressed MicroRNA-145,low-expressed MicroRNA-145,blank control vector,and transfect into HepG2 cells.Followed by retracting total RNA and protein to identify the expression level of ADD3 mRNA and ADD3 protein by using RT-PCR,qRT-PCR and werstern blot.The third part:detect the GBP1's expression in different degree of liver fibrosis,liver tissue,and using automatic medical image analysis system(HMIAS-2001 type)to quantify.The fourth part:build GBP1 gene lentivirus vectors,transfect liver cells so that raise its expression in liver cells,discuss its downstream factors especially the MMPs expression and the influence on the liver cells,when GBP 1 is high expression,and discussion the mechanism in liver fibrosis preliminary.Results:(1)The overexpression of ADD3 may be related to the pathogenesis of BA.(2)MicroRNA-145 may alter the expression of ADD3 by targeting the ADD3 3'UTR sequence and induce the occurrence of BA.(3)GBP 1 may be closely related with liver fibrosis in BA patients.(4)GBP 1 may participate in the development of BA by promoting liver cell apoptosis,destructing proliferation apoptosis balance and inhibiting the expression MMP 1,MMP-2 to decrease collagen enzyme hydrolysis.Since our study is just at a cellular level,further work is needed to be done in animal model.