Enhanced Expressions and Histological Characteristics of Intravenously Administered Plasmid DNA in Rat Lung.
10.3346/jkms.2001.16.5.567
- Author:
Suk Joo RHA
1
;
Young Pil WANG
Author Information
1. Department of Thoracic & Cardiovascular Surgery, College of Medicine, Catholic University of Korea, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
Gene Transfer Techniques;
Plasmid DNA;
Lung;
Transfection
- MeSH:
Animal;
DNA/*administration & dosage/metabolism;
Galactosides/analysis;
*Gene Therapy;
Gene Transfer, Horizontal;
Immunohistochemistry;
Indoles/analysis;
Injections, Intravenous;
Lung/*metabolism;
Male;
*Plasmids;
Rats;
Rats, Inbred F344;
*Transfection
- From:Journal of Korean Medical Science
2001;16(5):567-572
- CountryRepublic of Korea
- Language:English
-
Abstract:
Cationic liposome-mediated gene transfection is a promising method for gene therapy. In this study, the transfection efficiency and histological patterns were evaluated in rat lung after intravenous administration via femoral vein of naked plasmid DNA, naked plasmid DNA with pretreatment of DOTAP, and DOTAP-cholesterol-plasmid DNA complex. Plasmid DNA encoding bacterial LacZ gene was used. For quantification of LacZ gene expression, -galactosidase assay was performed. For histologic examination, X-gal staining and immunohistochemical staining for transfected gene products were performed. Pretreatment of DOTAP prior to the infusion of naked plasmid DNA increased transfection efficiency up to a level comparable to DOTAP-cholesterol-plasmid DNA complex injection. Transfected genes were mainly expressed in type II pneumocytes and alveolar macrophages in all animals. We conclude that the high transfection efficiency is achievable by intravenous administration of naked plasmid DNA with pretreatment of DOTAP, to a level comparable to DOTAP-cholesterol-plasmid DNA complex. In this regard, naked plasmid DNA administration with pretreatment of DOTAP could be a more feasible option for intravenous gene transfer than DOTAP-cholesterol-plasmid DNA complex, in that the former is technically easier and more cost-effective than the latter with a comparable efficacy, in terms of intravenous gene delivery to the lung.