Expression of interleukin 6 and apoptosis-related genes in suckling and weaning rat models of hepatectomy and liver regeneration.
OBJECTIVE
The most commonly used model to study the mechanisms of liver regeneration is the adult rat submitted to 70% to 80% hepatectomy. However, there are no studies using newborn or weaning rat models. The process of liver regeneration includes both the hypertrophy and hyperplasia of cells (processes regulated by growth factors and cytokines, mainly interleukin 6 [IL-6]) as well as apoptosis, or programmed cell death (a process regulated mainly by the Bcl-2 family of proteins). Proapoptotic proteins in this family include Bax and Bak. Conversely, Bcl-2 and Bcl-XL are antiapoptotic regulators. Therefore, to expand our understanding of liver regeneration, our study had 2 goals: first, to standardize 2 animal models of hepatectomy and liver regeneration using the newborn suckling and the weaning rat and second, to quantitate the expression levels of IL-6 and several members of the Bcl-2 gene family during the regeneration process.


METHODS
To create the experimental models, newborn suckling rats (age, 5-7 days; weight, 6-10 g) and weaning rats (age, 21-23 days; weight, 30-50 g) underwent 70% hepatectomy. The animals were subsequently sacrificed at days 1, 2, 3, 4, and 7 after hepatectomy, and the remnant liver lobes were harvested for routine histologic examination. Groups of healthy animals not operated on served as controls. For the experimental study, 6 newborn rats and 6 weaning rats underwent hepatectomy. The animals were killed 1 day after liver resection and the remnant livers were harvested to assess gene expression by quantitative reverse transcription-polymerase chain reaction. The hepatectomized groups were compared with control and sham groups.


RESULTS
During the creation of the experimental models, 70% of the suckling animals and all the weaning animals survived the hepatectomy. The decreased liver weight was completely restored to control levels by day 7 after hepatectomy. Histologically, the remnant livers of both hepatectomy groups exhibited steatosis, tumefaction of hepatocytes, and mitosis, which ceased at 7 days after the hepatectomy. The weaning rat model showed more robust gene expression responses. Specifically, expression levels of IL-6 gene were significantly increased after both surgical insult (sham group) and hepatectomy. However, this increase was significantly higher in the latter group. Furthermore, hepatectomy promoted a decrease in the expression levels of the proapoptotic genes and an increase in the expression levels of Bcl-2.


CONCLUSIONS
Our data indicate that regulation of both IL-6 and genes involved in apoptosis are strongly implicated in the mechanisms of liver regeneration and that the weaning rat model represents an attractive model system for future investigations in this area.
Coelho
Maria Cecília M
MC
Pediatric Surgery Laboratory, University of São Paulo Medical School, São Paulo 01246-903, Brazil.
Tannuri
Uenis
U
Tannuri
Ana Cristina A
AC
Mello
Evandro S
ES
dos Santos
Neide Aparecida S R
NA
D000818:Animals; D000833:Animals, Suckling; D017209:Apoptosis / Q000235:genetics*; D001835:Body Weight; D015870:Gene Expression; D019254:Genes, bcl-2 / Q000235:genetics; D006498:Hepatectomy; D015850:Interleukin-6 / Q000235:genetics* / Q000378:metabolism; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008115:Liver Regeneration / Q000235:genetics*; D023421:Models, Animal; D009929:Organ Size; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D051381:Rats; D014886:Weaning; D050998:bcl-2 Homologous Antagonist-Killer Protein / Q000235:genetics; D051028:bcl-2-Associated X Protein / Q000235:genetics; D051020:bcl-X Protein / Q000235:genetics
D015850:Interleukin-6; D019253:Proto-Oncogene Proteins c-bcl-2; D050998:bcl-2 Homologous Antagonist-Killer Protein; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein