Third-generation beta-blockers stimulate nitric oxide release from endothelial cells through ATP efflux: a novel mechanism for antihypertensive action.
BACKGROUND
Nebivolol and carvedilol are third-generation beta-adrenoreceptor antagonists, which unlike classic beta-blockers, have additional endothelium-dependent vasodilating properties specifically related to microcirculation by a molecular mechanism that still remains unclear. We hypothesized that nebivolol and carvedilol stimulate NO release from microvascular endothelial cells by extracellular ATP, which is a well-established potent autocrine and paracrine signaling factor modulating a variety of cellular functions through the activation of P2-purinoceptors.


RESULTS
Contraction and relaxation of renal glomerular vasculature were measured by determination of intracapillary volume with [3H]-inulin. Biologically active NO was measured with highly sensitive porphyrinic NO microsensors in a single glomerular endothelial cell (GEC). Extracellular ATP was measured by a luciferin-luciferase assay. Enzymatic degradation of extracellular ATP by apyrase and blockade of P2Y-purinoceptors by suramin or reactive blue 2 inhibited both beta-blocker-induced glomerular vasorelaxations and beta-blocker-stimulated NO release from GECs. Both beta-blocker-induced vasorelaxations were in the micromolar concentration range identical to that required for the beta-blocker stimulation of ATP and NO release from GECs. The maximum of NO release for nebivolol and carvedilol was very similar (188+/-14 and 226+/-17, respectively). Blockade of ATP release by a mechanosensitive ion channel blocker, Gd3+, inhibited the beta-blocker-dependent release of ATP and NO from GECs.


CONCLUSIONS
These results demonstrate for the first time that nebivolol and carvedilol induce relaxation of renal glomerular microvasculature through ATP efflux with consequent stimulation of P2Y-purinoceptor-mediated NO release from GECs.
Kalinowski
Leszek
L
Department of Chemistry and Biochemistry, Ohio University, Athens, Ohio 45701, USA.
Dobrucki
Lawrence W
LW
Szczepanska-Konkel
Miroslawa
M
Jankowski
Maciej
M
Martyniec
Ludmila
L
Angielski
Stefan
S
Malinski
Tadeusz
T
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000319:Adrenergic beta-Antagonists / Q000494:pharmacology*; D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D000959:Antihypertensive Agents / Q000494:pharmacology; D001578:Benzopyrans / Q000494:pharmacology; D002227:Carbazoles / Q000494:pharmacology; D000077261:Carvedilol; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D004983:Ethanolamines / Q000494:pharmacology; D005110:Extracellular Space / Q000378:metabolism; D005682:Gadolinium / Q000494:pharmacology; D007424:Intracellular Fluid / Q000378:metabolism; D007678:Kidney Glomerulus / Q000098:blood supply / Q000166:cytology; D008163:Luminescent Measurements; D008297:Male; D008833:Microcirculation / Q000166:cytology / Q000502:physiology; D000068577:Nebivolol; D009569:Nitric Oxide / Q000378:metabolism*; D011412:Propanolamines / Q000494:pharmacology; D051381:Rats; D018048:Receptors, Purinergic P2; D013498:Suramin / Q000494:pharmacology; D014661:Vasoconstriction / Q000187:drug effects / Q000502:physiology; D014664:Vasodilation / Q000187:drug effects / Q000502:physiology
D000319:Adrenergic beta-Antagonists; D000959:Antihypertensive Agents; D001578:Benzopyrans; D002227:Carbazoles; D004983:Ethanolamines; D011412:Propanolamines; D018048:Receptors, Purinergic P2; D000068577:Nebivolol; D000077261:Carvedilol; D000804:Angiotensin II; D009569:Nitric Oxide; D013498:Suramin; D000255:Adenosine Triphosphate; D005682:Gadolinium