| Objective: With the rapid improvement of the molecular biological theories and recombinant DNA techniques, specifically, the gene transfer techniques in genetic engineering, modifications of donated organs before implantation may improve graft function and overall survival to prevent or attenuate complications such as ischemia-reperfusion injury, rejection and infection. This study is to determine the feasibility of ex vivo liposome-mediated gene transfer ( Lac-Z gene ) to lung isografts and to get expression. Methods: An orthotopic rat left lung transplant model was developed with the use of a modification of the "cuff technique. 18 SD rats ( female for donor, male for recipient ) were divided into three groups randomly. Gene transfer group ( group 1 ): PSV- (3 -gal/ liposome ( Lipofectin8 ) complex was transfered into the donor left lung via the left pulmonary vascular route ( via the left pulmonary vein route ) at the time of lung harvesting. Liposome group ( group 2 ) and Control group ( group 3 ) : Grafts underwent the same procedure but received liposome and 0.9% saline solution respectively. Donor lungs were harvested one day after transplantation. The transgene expression of Lac-Z gene was detected by histochemical staining. Results: Transgene expression of P -gal in group 1 was detected in bronchial epithelial cells and alveolar epithelial cells. But no expression of 3 -gal was found in group 2 and 3. Conclusion: Ex vivo plasmid vector/liposome complex-mediated gene transfer to lung isografts via the left pulmonary vein route is feasible. This study provides the scientific proof to the feasibility of functional gene therapy of complications such as ischemia-reperfusion injury, rejection and infection after rat lung transplantation.
|
PDF Full Text Request