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.
In pediatric liver transplantations with LFS grafts, higher incidences of graft dysfunction probably occur due to IRI. It was postulated that increasing the blood supply to the graft by means of a meso-caval shunt could ameliorate the IRI. Eleven pigs underwent liver transplantation and were divided into two groups: LFS and LFS+SHUNT group. A series of flowmetric, metabolic, histologic, and molecular studies were performed. No significant metabolic differences were observed between the groups. One hour after reperfusion, portal flow was significantly lower in the recipients than in the donors, proving that the graft was maintained in low portal blood flow, although the shunt could promote a transient increase in the portal blood flow and a decrease in the arterial flow. Finally, it was verified that the shunt promoted a decrease in inflammation and steatosis scores and a decrease in the expression of the eNOS gene (responsible for the generation of nitric oxide in the vascular endothelium) and an increase in the expression of the proapoptotic gene BAX. The meso-caval shunt was responsible for some positive effects, although other deleterious flowmetric and molecular alterations also occurred.
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