Activation of AMP kinase alpha1 subunit induces aortic vasorelaxation in mice.
Vasodilatation is a vital mechanism of systemic blood flow regulation that occurs during periods of increased energy demand. The AMP-dependent protein kinase (AMPK) is a serine/threonine kinase that is activated by conditions that increase the AMP-to-ATP ratio, such as exercise and metabolic stress. We hypothesized that AMPK could trigger vasodilatation and participate in blood flow regulation. Rings of thoracic aorta were isolated from C57Bl6 mice and mice deficient in the AMPK catalytic alpha1 (AMPKalpha1-/-) or alpha2 (AMPKalpha2-/-) subunit and their littermate controls, and mounted in an organ bath. Aortas were preconstricted with phenylephrine (1 microM) and activation of AMPK was induced by addition of increasing concentrations of 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR). AICAR (0.1-3 mM) dose-dependently induced relaxation of precontracted C57BL6, AMPKalpha1+/+ and alpha2+/+ aorta (P<0.001, n=5-7 per group). This AICAR induced vasorelaxation was not inhibited by the addition of adenosine receptor antagonists. Moreover, when aortic rings were freed of endothelium by gentle rubbing, AICAR still induced aortic ring relaxation, suggesting a direct effect of AICAR on smooth muscle cells. When aortic rings were pretreated with L-NMMA (30 microM) to inhibit nitric oxide synthase activity, AICAR still induced relaxation. Western blot analysis of C57Bl6 mice denuded aorta showed that AMPK was phosphorylated after incubation with AICAR and that AMPKalpha1 was the main catalytic subunit expressed. Finally, AICAR-induced relaxation of aortic rings was completely abolished in AMPKalpha1-/- but not AMPKalpha2-/- mice. Taken together, the results show that activation of AMPKalpha1 but not AMPKalpha2 is able to induce aortic relaxation in mice, in an endothelium- and eNOS-independent manner.
Goirand
Françoise
F
Université Paris-Sud, IFR-141, Châtenay-Malabry, France. goirand.francoise@wanadoo.fr
Solar
Myriam
M
Athea
Yoni
Y
Viollet
Benoit
B
Mateo
Philippe
P
Fortin
Dominique
D
Leclerc
Jocelyne
J
Hoerter
Jacqueline
J
Ventura-Clapier
Renée
R
Garnier
Anne
A
D055372:AMP-Activated Protein Kinases; D000620:Aminoimidazole Carboxamide / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects / Q000201:enzymology*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D020536:Enzyme Activators / Q000494:pharmacology; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009097:Multienzyme Complexes / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000201:enzymology; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D012265:Ribonucleotides / Q000494:pharmacology; D014664:Vasodilation / Q000187:drug effects
D020536:Enzyme Activators; D007527:Isoenzymes; D009097:Multienzyme Complexes; D012265:Ribonucleotides; D000620:Aminoimidazole Carboxamide; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases; C031143:AICA ribonucleotide