Planta Med 2005; 71(9): 829-834
DOI: 10.1055/s-2005-871296
Original Paper
Pharmacology
© Georg Thieme Verlag KG Stuttgart · New York

Endothelial Nitric Oxide Production Stimulated by the Bioflavonoid Chrysin in Rat Isolated Aorta

Inmaculada Concepción Villar1 , 3 , Rocío Vera1 , 3 , Milagros Galisteo1 , Francisco O’Valle2 , Miguel Romero1 , Antonio Zarzuelo1 , Juan Duarte1
  • 1Departamento de Farmacología, Facultad de Farmacia, Universidad de Granada, Granada, Spain
  • 2Departamento de Patología, Facultad de Medicina, Universidad de Granada, Granada, Spain
  • 3I. C. Villar and R. Vera contributed equally to this work
Further Information

Publication History

Received: November 11, 2004

Accepted: April 25, 2005

Publication Date:
19 August 2005 (online)

Abstract

In the present study, the effects of the bioflavonoid chrysin (5,7-dihydroxyflavone) were analysed on nitric oxide (NO) production from vascular endothelium. In aortic rings, incubation with chrysin or acetylcholine (both at 10 μM) increased L-NAME-sensitive endothelial NO release as measured using the fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2 DA). Moreover, chrysin increased cGMP accumulation only in aortic rings with endothelium. However, at this concentration, chrysin had no effect either on basal or on NADPH-stimulated vascular superoxide production. Moreover, at this low concentration, chrysin, similar to acetylcholine, induced aortic relaxation, which was abolished by both endothelial deprivation and NO synthase inhibition. Endothelium-dependent relaxation induced by chrysin was unaltered by removal of extracellular calcium and incubation with the intracellular calcium chelator BAPTA, while the phosphatidylinositol (PI)-3 kinase inhibitor wortmannin suppressed the endothelial dependence. In conclusion, chrysin stimulated NO release from endothelial cells leading to vascular cGMP accumulation and subsequent endothelium dependent aortic relaxation. Chrysin-stimulated NO release is calcium independent and possibly mediated via PI3-kinase.

References

  • 1 Kühnau J. The flavonoids. A class of semi-essential food components: their role in human nutrition.  World Rev Nutr Diet. 1976;  24 117-91
  • 2 Chan E CH, Pannangpetch P, Woodman O L. Relaxation to flavones and flavonols in rat isolated thoracic aorta: mechanism of action and structure-activity relationships.  J Cardiovasc Pharmacol. 2000;  35 326-33
  • 3 Duarte J, Pérez-Vizcaino F, Utrilla M P, Jiménez J, Tamargo J, Zarzuelo A. Vasodilating effects of flavonoids in rat aortic smooth muscle: structure-activity relationships.  Gen Pharmacol. 1993;  24 857-62
  • 4 Herrera M D, Zarzuelo A, Jimenez J, Marhuenda E, Duarte J. Effects of flavonoids on rat aortic smooth muscle contractility: structure-activity relationships.  Gen Pharmacol. 1996;  27 273-7
  • 5 Ajay M, Gilani A U, Mustafa M R. Effects of flavonoids on vascular smooth muscle of the isolated rat thoracic aorta.  Life Sci. 2003;  74 603-12
  • 6 Beretz A, Stoclet J C, Anton R. Inhibition of isolated rat aorta contraction by flavonoids. Possible correlation with cAMP-phosphodiesterase inhibition.  Planta Med. 1980;  39 236
  • 7 Perez-Vizcaino F, Ibarra M, Cogolludo A L, Duarte J, Zaragoza-Arnaez F, Moreno L. et al . Endothelium-independent vasodilator effects of the flavonoid quercetin and its methylated metabolites in rat conductance and resistance arteries.  J Pharmacol Exp Ther. 2002;  302 66-72
  • 8 Siess M H, Le Bon A M, Canivenc-Lavier M C, Amiot M J, Sabatier S, Aubert S Y. et al . Flavonoids of honey and propolis: Characterization and effects on hepatic drug-metabolizing enzymes and benzo[a]pyrene-DNA binding in rats.  J Agric Food Chem. 1996;  44 2297-301
  • 9 Hertog M GL, Hollman P CH, Katan M B. Content of potentially anticarcinogenic flavonoids of 28 vegetables and 9 fruits commonly consumed in the Netherlands.  J Agric Food Chem. 1992;  40 2379-83
  • 10 Duarte J, Jiménez R, Villar I C, Pérez-Vizcaíno F, Jiménez J, Tamargo J. Vasorelaxant effects of the bioflavonoid chrysin in isolated rat aorta.  Planta Med. 2001;  67 567-9
  • 11 Villar I C, Galisteo M, Vera R, O’Valle F, Garcia-Saura M F, Zarzuelo A. et al . Effects of the dietary flavonoid chrysin in isolated rat mesenteric vascular bed.  J Vasc Res. 2004;  41 509-16
  • 12 Maffei A, Poulet R, Vecchione C, Colella S, Fratta L, Frati G. et al . Increased basal nitric oxide release despite enhanced free radicals production in hypertension.  J Hypertens. 2002;  20 1135-42
  • 13 Ohara Y, Peterson T E, Harrison D G. Hypercholesterolemia increases endothelial superoxide anion production.  J Clin Invest. 1993;  91 2546-51
  • 14 Vanhoutte P M, Rubanyi G M, Miller V M, Houston D S. Modulation of vascular smooth muscle contraction by the endothelium.  Annu Rev Physiol. 1986;  48 307-20
  • 15 Warner T D, Mitchell J A, Sheng H, Murad F. Effects of cyclic GMP on smooth muscle relaxation.  Adv Pharmacol. 1994;  26 171-94
  • 16 Carr A, Frei B. The role of natural antioxidants in preserving the biological activity of endothelium-derived nitric oxide.  Free Radic Biol Med. 2000;  28 1806-14
  • 17 Mohazzab K M, Kaminski P M, Wolin M S. NADH oxidoreductase is a major source of superoxide anion in bovine coronary artery endothelium.  Am J Physiol. 1994;  266 H2568-72
  • 18 Rajagopalan S, Kurz S, Munzel T, Tarpey M, Freeman B A, Griendling K K. et al . Angiotensin II-mediated hypertension in the rat increases vascular superoxide production via membrane NADH/NADPH oxidase activation. Contribution to alterations of vasomotor tone.  J Clin Invest. 1996;  97 1916-23
  • 19 Gryglewski R J, Palmer R M, Moncada S. Superoxide anion is involved in the breakdown of endothelium-derived vascular relaxing factor.  Nature. 1986;  320 454-6
  • 20 Grunfeld S, Hamilton C A, Mesaros S, McClain S W, Dominiczak A F, Bohr D F. et al . Role of superoxide in the depressed nitric oxide production by the endothelium of genetically hypertensive rats.  Hypertension. 1995;  26 854-7
  • 21 Tschudi M R, Mesaros S, Luscher T F, Malinski T. Direct in situ measurement of nitric oxide in mesenteric resistance arteries. Increased decomposition by superoxide in hypertension.  Hypertension. 1996;  27 32-5
  • 22 Stuehr D J. Structure-function aspects in the nitric oxide synthases.  Annu Rev Pharmacol Toxicol. 1997;  37 339-59
  • 23 Hartell N A, Archer H E, Bailey C J. Insulin-stimulated endothelial nitric oxide release is calcium independent and mediated via protein kinase B.  Biochem Pharmacol. 2005;  69 781-90

J. Duarte

Departamento de Farmacología

Facultad de Farmacia

Universidad de Granada

18071 Granada

Spain

Phone: +34-958-244-088

Fax: +34-958-248-964

Email: jmduarte@platon.ugr.es

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