Protein kinase C masks nitric oxide synthase activity in vascular smooth muscle under basal conditions.
Under basal conditions there is no observable nitric oxide synthase (NOS) activity in vascular smooth muscle (VSM). Pretreatment of endothelium-denuded aortic rings from Sprague-Dawley rats with 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7), (0.1 micromol/L) significantly attenuated phenylephrine (PE)-induced contractile responses in a dose-dependent manner. In the presence of 10 micromol/L Nomega-nitro-L-arginine (L-NNA) or 0.1 mmol/L aminoguanidine (AG), the inhibition of contractions at 10 nmol/L PE by H-7 was blocked by 88% or 52%, respectively. The blockade by antagonists was completely reversed by l-arginine but not by d-arginine, and alone they did not significantly alter PE-induced contraction of endothelium-denuded aorta. Methylene blue (MB, 50 micromol/L) also inhibited the action of H-7. The inhibitory effect of H-7 occurred after 5 minutes and was reversible. PE-induced contraction was also inhibited by the selective protein kinase C inhibitors calphostin C (10 micromol/L), and bisindolylmaleimide IV (Bis-IV, 10 micromol/L), but not by the selective protein kinase A inhibitor H-89 (0.1 micromol/L). These results indicate protein kinase C inhibits NOS activity in VSM under basal conditions. Incubation of tissues with either H-7 or calphostin C stimulates NO production, and immunocytochemical studies reveal the presence of NOS in VSM under basal conditions.
Kline
Loren W
LW
Department of Physiology, University of Alberta, Edmonton, Alberta, Canada.
Ji
Junzhi
J
Wang
Guei-Jane
GJ
Sutherland
Sharla K
SK
Pang
Peter K T
PK
Benishin
Christina G
CG
D000818:Animals; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D019335:Nitroarginine / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014661:Vasoconstriction / Q000187:drug effects
D004791:Enzyme Inhibitors; D019335:Nitroarginine; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D011493:Protein Kinase C