Effects and mechanism of adenovirus-mediated phosphatase and tension homologue deleted on chromosome ten gene on collagen deposition in rat liver fibrosis.

AIM

To evaluate the effects of phosphatase and tension homologue deleted on chromosome ten () gene on collagen metabolism in hepatic fibrosis and the underlying mechanisms.

METHODS

Rat primary hepatic stellate cells (HSCs) and human LX-2 cells were transfected with adenovirus containing cDNA constructs encoding wild-type (Ad-PTEN), mutant gene (Ad-G129E), and RNA interference constructs targeting the sequence short hairpin RNA to up-regulate and down-regulate the expression of . HSCs were assayed using fluorescent microscopy, real-time polymerase chain reaction, and western blotting. Moreover, a CCl-induced rat hepatic fibrosis model was established to investigate the effects. Hematoxylin and eosin, and Masson's trichrome were used to assess the histological changes. The expression of collagen I and III was assessed using immunohistochemistry and western blot analysis.

RESULTS

Elevated expression of gene reduced serum levels of alanine transaminase and aspartate transaminase, decreased collagen deposition in the liver, and reduced hepatocyte necrosis. In contrast, knockdown of expression had an opposite effect, such as increased collagen deposition in the liver, and was molecularly characterized by the increased expression of matrix metalloproteinase (MMP)-13 ( < 0.01) and MMP-2 ( < 0.01), as well as decreased expression of the tissue inhibitor of metalloproteinase (TIMP)-1 ( < 0.01) and TIMP-2 ( < 0.01).

CONCLUSION

These data indicated that gene therapy using recombinant adenovirus encoding PTEN might be a novel way of treating hepatic fibrosis.