Vascular inflammation inhibits gene transfer to the pulmonary circulation in vivo
| Autor: | Robert C. Tyler, Robert C. Unfer, Cornelia Gorman, Molly McClarrion, David M. Rodman, Karen A. Fagan, Clayton M. Bullock |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Chloramphenicol O-Acetyltransferase
Vasculitis Pulmonary and Respiratory Medicine Pulmonary Circulation Pathology medicine.medical_specialty Physiology Genetic enhancement Gene Expression Mice Transgenic Inflammation Pharmacology Transfection Mice Genes Reporter In vivo Physiology (medical) medicine Animals RNA Messenger Transgenes Respiratory system Mice Inbred ICR Liposome Monocrotaline business.industry Genetic transfer Respiratory disease Cell Biology medicine.disease Pulmonary hypertension Blotting Southern Liposomes Female medicine.symptom business Plasmids |
| Zdroj: | American Journal of Physiology-Lung Cellular and Molecular Physiology. 277:L1199-L1204 |
| ISSN: | 1522-1504 1040-0605 |
| DOI: | 10.1152/ajplung.1999.277.6.l1199 |
| Popis: | We report gene transfer to the normal and injured murine pulmonary circulation via systemic (intravascular) and airway (intratracheal) delivery of novel polycationic liposomes (imidazolium chloride, imidazolinium chloride-cholesterol, and ethyl phosphocholine). With use of the reporter genes chloramphenicol acetyltransferase ( CAT) or human placental alkaline phosphatase ( hpAP), intravascular injection of lipid-DNA complexes resulted in gene expression primarily in the lung, with lesser expression in the heart (11% of lung, P < 0.05) and spleen (8% of lung, P < 0.05). Histochemical staining for the hpAPreporter gene showed localized transgene expression in the microvascular endothelium. Monocrotaline (80 mg/kg body wt sc) treatment produced endovascular inflammation and reduced lung CAT activity (2 days postintravascular transfection) by 75 ± 8 and 86 ± 6% at 7 and 21 days, respectively, after monocrotaline ( P < 0.05). Despite the apparent decrease in functional CAT protein, Southern blot analysis suggested maintained plasmid delivery, whereas quantitative PCR ( TaqMan) showed decreased CAT mRNA levels in monocrotaline mice. In contrast, intratracheal delivery of lipid-DNA complexes showed enhanced CAT expression in monocrotaline mice. Transfection in alternate pulmonary vascular disorders was studied by inducing hypoxic pulmonary hypertension (4 wk at barometric pressure of 410 mmHg). Efficiency and duration of gene transfer, assessed by CAT activity, were similar in pulmonary hypertensive and normal lungs. We conclude that imidazolinium-derived polycationic liposomes provide a means of relatively selective and efficient gene transfer to the normal and injured murine microvascular circulation, although translation of transgene mRNA may be reduced by preexisting endothelial injury. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|