Application of hydrostatic pressure facilitates ex vivo adenovirus gene transfer into rabbit aortas.
We investigated whether application of hydrostatic pressure facilitates ex vivo adenovirus gene transfer into rabbit aortas. Assay of beta-galactosidase activity was performed in the aortas subjected to adenovirus encoding LacZ gene (Ad-LacZ) transfer. Application of hydrostatic pressure at 8 atmospheres during Ad-LacZ gene transfer (10(10) pfu/mL, 10 min) to aortic segments resulted in an approximately 4.5-fold increase in transgene efficiency. X-Gal staining showed predominant beta-galactosidase activity in the endothelial and the adventitial cells in the aorta subjected to pressure-supported Ad-LacZ gene transfer. Then we examined the effect of heme oxygenase-1 (HO-1), which catabolizes heme to carbon monoxide and biliverdin, by transferring adenovirus encoding HO-1 (Ad-HO-1) into rabbit aortas using this pressurization system. The Ad-HO-1-infected aortic segment showed a significantly decreased contractile response to phenylephrine compared to the Ad-LacZ-infected aortic segment. Pressure-supported adenovirus gene transfer may increase the feasibility of exploiting intraoperative adenovirus-mediated gene transfer.
Department of Cardiovascular Medicine, University of Tokyo Graduate School of Medicine, Hongo 7-3-1, Bunkyo-ku, Tokyo, 113-8655, Japan.
M
M
Suzuki
N
N
Ishizaka
W M
WM
Hwang
K
K
Tsukamoto
K
K
Minami
T
T
Aizawa
I
I
Mori
J
J
Taguchi
R
R
Nagai
M
M
Ohno
D000109:Acetylcholine / Q000494:pharmacology; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001011:Aorta / Q000166:cytology / Q000378:metabolism*; D015153:Blotting, Western; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005697:Galactosides / Q000378:metabolism; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006419:Heme Oxygenase (Decyclizing) / Q000037:antagonists & inhibitors / Q000235:genetics; D051547:Heme Oxygenase-1; D006874:Hydrostatic Pressure; D066298:In Vitro Techniques; D007211:Indoles / Q000378:metabolism; D008297:Male; D008665:Metalloporphyrins / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010656:Phenylephrine / Q000494:pharmacology; D011524:Protoporphyrins / Q000494:pharmacology; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D014661:Vasoconstriction / Q000187:drug effects; D014662:Vasoconstrictor Agents / Q000494:pharmacology; D001616:beta-Galactosidase / Q000235:genetics
D004791:Enzyme Inhibitors; D005697:Galactosides; D007211:Indoles; D008665:Metalloporphyrins; D011524:Protoporphyrins; D014662:Vasoconstrictor Agents; D010656:Phenylephrine; C032628:tin protoporphyrin IX; D006419:Heme Oxygenase (Decyclizing); D051547:Heme Oxygenase-1; D001616:beta-Galactosidase; D000109:Acetylcholine; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside