Adeno-associated virus-mediated transfer of endothelial nitric oxide synthase gene inhibits protein synthesis of rat ventricular cardiomyocytes.
We investigated whether nitric oxide (NO) synthase gene transfer could attenuate growth of cultured cardiac myocytes. First, we investigated the effects of exogenous NO and cGMP analog on protein synthesis of cultured neonatal rat cardiac myocytes. The NO donor 3-morpholino-sydnonimine-hydrochloride (SIN-1) and 8-bromo-cGMP caused concentration-dependent decreases in phenylephrine-stimulated incorporation of 3H-leucine into cardiac myocytes. We then transferred endothelial constitutive NO synthase (ecNOS) gene into cultured neonatal rat cardiac myocytes using adeno-associated virus (AAV) vectors. ecNOS gene transfer into cardiac myocytes induced 140 kD ecNOS protein expression and significantly increased cGMP contents of myocytes compared with control cells. ecNOS gene transfer inhibited 3H-leucine incorporation into cardiac myocytes in response to phenylephrine, which was significantly recovered in the presence of the NOS inhibitor N(G)-monomethyl-L-arginine acetate. These results indicate that endogenously generated NO by ecNOS gene transfer using AAV vectors inhibits the alpha-adrenergic agonist-induced cardiac protein synthesis at least partially via cGMP production.
Maeda
Y
Y
Department of Cardiology, Jichi Medical School, Tochigi, Japan.
Ikeda
U
U
Oya
K
K
Shimpo
M
M
Ueno
S
S
Urabe
M
M
Kume
A
A
Monahan
J
J
Ozawa
K
K
Shimada
K
K
D000256:Adenoviridae / Q000235:genetics*; D000316:Adrenergic alpha-Agonists / Q000494:pharmacology; D000818:Animals; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005822:Genetic Vectors; D006352:Heart Ventricles / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015151:Immunoblotting; D007930:Leucine / Q000378:metabolism; D009124:Muscle Proteins / Q000096:biosynthesis*; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000235:genetics*; D052250:Nitric Oxide Synthase Type III; D010656:Phenylephrine / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D051381:Rats; D019323:omega-N-Methylarginine / Q000494:pharmacology
D000316:Adrenergic alpha-Agonists; D004791:Enzyme Inhibitors; D009124:Muscle Proteins; D010656:Phenylephrine; D019323:omega-N-Methylarginine; C016276:8-bromocyclic GMP; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat; D007930:Leucine; D006152:Cyclic GMP