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Hydrodynamics based transfection in normal and fibrotic rats

World Journal of Gastroenterology 2006年第38期12卷 6149-6155页

作者： Rita Yeikilis Shunit gal.Natalia Kopeiko Melia Paizi Mark Pines Filip Braet Gadi Spira Department of Anatomy and Cell Biology The Bruce Rappaport Faculty of Medicine Rappaport Family Institute for Research in the Medical Sciences Technion Haifa 31096 Israel Department of Experimental Surgery The Bruce Rappaport Faculty of Medicine Rappaport Family Institute for Research in the Medical Sciences Technion Haifa 31096 Israel Institute of Animal Sciences Agricultural Research Organization the Volcani Center Bet Dagan 50250 Israel Australian Key Centre for Microscopy and Microanalysis The University of Sydney Sydney NSW 2006 Australia

AIM： Hydrodynamics based transfection （HBT）, the injection of a large volume of naked plasmid DNA in a short time is a relatively simple, efficient and safe method for in vivo transfection of liver cells. Though used for quite some time, the mechanism of gene transfection has not yet been elucidated. METHODS： A lucJferase encoding plasmid was injected using the hydrodynamics based procedure into normal and thioacetamide-induced fibrotic Sprague Dawley rats. Scanning and transmission electron microscopy images were taken. The consequence of a dual injection of Ringer solution and luciferase pDNA was followed. Halofuginone, an anti collagen type I inhibitor was used to reduce ECM load in fibrotic rats prior to the hydrodynamic injection. RESULTS： Large endothelial gaps formed as soon as 10＇ following hydrodynamic injection; these gradually returned to normal 10 d post injection. Hydrodynamic administration of Ringer 10 or 30 m prior to moderate injection of plasmid did not result in efficient transfection suggesting that endothelial gaps by themselves are not sufficient for gene expression. Gene transfection following hydrodynamic injection in thioacetamide induced fibrotic rats was diminished coinciding with the level of fibrosis. Halofuginone, a specific collagen type I inhibitor, alleviated this effect. CONCLUSION： The hydrodynamic pressure formed following HBT results in the formation of large endothelial gaps. These gaps, though important in the transfer of DNA molecules from the blood to the space of Disse are not enough to provide the appropriate conditions for hepatocyte transfection. Hydrodynamics based injection is applicable in fibrotic rats provided that ECM load is reduced.

关键词： Gene transfection Fibrosis In vivo transfection Fibrotic Hepatic Endothelial lining Endothelium Sinusoidal Fenestrae Space of disse Extracellular matrix

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