Figure 3
Time course for experimental testing of antifibrotic devices directed against TGF-β. (A) Male Sprague-Dawley rats were given a single infusion of either Ad5-CMV-AS-TGF-β1, Ad5-CMV-EGFP, or phosphate buffered saline at day 0 and a replenishment infusion at day 7. Two days after the first injection, the animals were subjected to bile duct ligation (BDL). At day 12, animals were sacrificed for liver histology and sampling of protein and RNA specimens. (B) Transfer of adenoviruses into normal and injured livers. A reporter adenovirus expressing the green fluorescent protein (EGFP) was injected two days prior surgery via the tail vein. Four days after surgery, the livers were homogenized, and 20 μg of protein samples each were tested for expression of EGFP by Western blot analysis. Note the lower level of transgene expression in animals that received bile duct ligature. (C) Transduction of AS-TGF-β1 in normal and injured livers in vivo. Liver injury was induced by BDL and adenoviruses (1 × 1010 pfu/kg) were administered to normal or injured rats two days prior BDL. Liver samples were analyzed for expression of the transgene by RT-PCR.