title: Dynamic expression of desmin, α-SMA and TGF-β1 during hepatic fibrogenesis induced by selective bile duct ligation in young rats.
abstract: We previously described a selective bile duct ligation model to elucidate the process of hepatic fibrogenesis in children with biliary atresia or intrahepatic biliary stenosis. Using this model, we identified changes in the expression of alpha smooth muscle actin (α-SMA) both in the obstructed parenchyma and in the hepatic parenchyma adjacent to the obstruction. However, the expression profiles of desmin and TGF-β1, molecules known to be involved in hepatic fibrogenesis, were unchanged when analyzed by semiquantitative polymerase chain reaction (RT-PCR). Thus, the molecular mechanisms involved in the modulation of liver fibrosis in this experimental model are not fully understood. This study aimed to evaluate the molecular changes in an experimental model of selective bile duct ligation and to compare the gene expression changes observed in RT-PCR and in real-time quantitative PCR (qRT-PCR). Twenty-eight Wistar rats of both sexes and weaning age (21-23 days old) were used. The rats were separated into groups that were assessed 7 or 60 days after selective biliary duct ligation. The expression of desmin, α-SMA and TGF-β1 was examined in tissue from hepatic parenchyma with biliary obstruction (BO) and in hepatic parenchyma without biliary obstruction (WBO), using RT-PCR and qRT-PCR. The results obtained in this study using these two methods were significantly different. The BO parenchyma had a more severe fibrogenic reaction, with increased α-SMA and TGF-β1 expression after 7 days. The WBO parenchyma presented a later, fibrotic response, with increased desmin expression 7 days after surgery and increased α-SMA 60 days after surgery. The qRT-PCR technique was more sensitive to expression changes than the semiquantitative method.
authors.affiliation: Laboratório de Pesquisa em Cirurgia Pediátrica (LIM-30), Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil.
authors.forename: J O
authors.initials: JO
authors.lastname: Gonçalves
authors.affiliation: Laboratório de Pesquisa em Cirurgia Pediátrica (LIM-30), Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil.
authors.forename: A C A
authors.initials: AC
authors.lastname: Tannuri
authors.affiliation: Laboratório de Pesquisa em Cirurgia Pediátrica (LIM-30), Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil.
authors.forename: M C M
authors.initials: MC
authors.lastname: Coelho
authors.affiliation: Laboratório de Pesquisa em Cirurgia Pediátrica (LIM-30), Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil.
authors.forename: I
authors.initials: I
authors.lastname: Bendit
authors.affiliation: Laboratório de Pesquisa em Cirurgia Pediátrica (LIM-30), Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil.
authors.forename: U
authors.initials: U
authors.lastname: Tannuri
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism*; D000704:Analysis of Variance; D000818:Animals; D001652:Bile Ducts / Q000601:surgery; D001656:Biliary Atresia; D002779:Cholestasis / Q000150:complications*; D024042:Collagen Type I / Q000096:biosynthesis; D003893:Desmin / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D005260:Female; D015870:Gene Expression; D008026:Ligation; D008099:Liver / Q000378:metabolism* / Q000601:surgery; D008103:Liver Cirrhosis / Q000209:etiology* / Q000378:metabolism; D008297:Male; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; D024042:Collagen Type I; D003893:Desmin; D053773:Transforming Growth Factor beta1; C498862:Acta2 protein, rat