Metallothionein 단백질 전달을 이용한 췌장소도 생존 향상 기술 연구

Abstract

Subcutaneous site is ideal for clinical islet transplantation because it has the advantage of simple operation procedure under local anesthesia and can be biopsied when needed. But, the transplantation outcomes at subcutaneous site have been disappointing due to oxidative stress induced by poor vascularization. We hypothesized that islet grafts would be subcutaneously transplanted using anti-oxidant enzyme metallothionein (MT) delivered to pancreatic islets by Tat protein transduction domain-mediated transduction. The Tat peptide-mediated MT delivery was confirmed by confocal microscopy, TEM and FACS analysis. The intracellular transduction of Tat peptide fused MT was found to increase in a dose-dependent manner, when various concentrations of the fusion protein were treated to islet. Once inside the cells, transduced Tat-MT protein was biologically active and showed protection effect under oxidative stress induced by intracellular nitric oxide donor, sodium nitroprusside. Furthermore, the cell viability and insulin secretion capability was not damaged by Tat-MT fusion protein treatment. Collectively, these results suggest that Tat-MT delivery into islet can offer a new strategy for clinically transplanted islets into subcutaneous site.