Expression, Reinigung und therapeutischer Einsatz eines löslichen TGF-[beta]-Typ-II-Rezeptors in der experimentell induzierten Leberfibrose
The aim of this work was the production and testing of a soluble tgf-beta type II receptor as a pure protein in the therapy of the experimental induced liverfibrosis. Tgf-beta is as a mastercytocine responsible for the transdifferentiation of hepatic stellate cells to myofiborblasts, the key-event of liverfibrosis. The protein may intercept tgf-beta before binding the hepatic stellate cells. For amplification of the protein COS-7 cells were infected with the adenoviral construct Ad5-CMV-sTbetaRII. The supernatant of the cells was purified with the HiTrap Protein G HP 1 ml column. By using 4x10exp11 pfu virus an amount of 1 g pure protein was harvested. The biological effect was proved by a PAI-1/luciferase-assay. In vitro testing happened by transdifferentiation assay with activated hepatic stellate cells. In vivo testing was performed by bile-duct-ligation induced liverfibrosis in male sprague dawley rats (mean bodyweight 250 g). Over a time-period of 14 days 0.1 mg/kg of the pure protein were administered by one intravenous and four intraperitoneal injections. For collagen I was a significant suppression here in northern-blot shown in animals treated with the protein in contrast to the control-group. In histologic-measurement there was a reduction of approximately five percent in the treated group.
info:eu-repo/classification/ddc/610, Medizin, Leberfibrose, TGF-beta, löslicher TGF-beta Typ II Rezeptor, Hepatische Stern Zellen, Myofibroblasten, liverfibrosis, soluble tgf-beta type II receptor, hepatic stellate cells, myofibroblasts
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