Date of Award

2010

Degree Type

Thesis

Degree Name

Master of Science

Program

Pathology

Supervisor

Dr. Madeleine Moussa

Second Advisor

Dr. Weiping Min

Abstract

TGF-βl is a pluripotent cytokine with multifunctional properties depending on the target cell type and origin. It has been known to play an important role in tissue repair and fibrosis. TGF-βl blockage has been studied over the years in order to treat fibrotic diseases, however; it resulted in inflammation as TGF-βl is also an anti-inflammatory cytokine. Normal Rat Kidney fibroblasts (NRK 49F) have been commonly used to study the fibrotic effects of TGF-βl. In this study, Smad3, a known key mediator of TGF-βl induced fibrosis, was silenced in NRK 49F cells and the silencing outcomes were studied in sets of in-vitro models. Smad3 expression, at mRNA levels, showed 85-90% silencing using Smad3 siRNA. Smad3 siRNA silenced cells showed less nodule formation, with regards to nodule size and the formation time compared to control, when stimulated with TGF-βl. Moreover, in the in-vitro wound healing model used, the Smad3 siRNA silenced cells showed less migration capabilities compared to control when stimulated with TGF-βl. Therefore, silencing Smad3 may be a therapeutic candidate for the treatment of fibrotic diseases and perhaps in transplantation.

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