title: Quercetin-induced induction of the NO/cGMP pathway depends on Ca2+-activated K+ channel-induced hyperpolarization-mediated Ca2+-entry into cultured human endothelial cells.
abstract: Quercetin is one of the dietary-derived flavonoids that are held responsible for the beneficial effects of red wine drinking in coronary artery disease known as the "French paradox". We examined whether quercetin modulates endothelial function by influencing Ca2+-activated K+ channels with large conductance (BK(Ca)) in cultured human endothelial cells. Membrane potential and intracellular Ca2+ concentrations of cultured human endothelial cells derived from umbilical cord veins (HUVEC) were measured using the fluorescence dyes DiBAC, and FURA-2, respectively. NO production was examined using a cGMP radioimmunoassay. HUVEC proliferation was analyzed by cell counts and thymidine incorporation. A dose-dependent hyperpolarization of HUVEC was recorded when quercetin was added (5-100 micromol/L). The maximum effect (50 micromol/L) was significantly reduced by the addition of the highly selective BK(Ca) inhibitor iberiotoxin (100 nmol/L), but not by blockers of other Ca2+-activated K+ channels (n = 30; p < 0.05). This BK(Ca)-induced hyperpolarization caused a transmembrane Ca2+ influx, because the quercetin-induced increase of intracellular Ca2+ was blocked by iberiotoxin, or by applying 2-aminoethoxydiphenylborate (100 micromol/L)--an inhibitor of capacitative Ca2+ entry (n = 30; p < 0.05). Quercetin-induced cGMP levels were significantly reduced by the eNOS-inhibitor l-NMMA (300 micromol/L), and by iberiotoxin (n = 10; p < 0.05). Endothelial proliferation was significantly reduced by 56 % when cells were incubated with quercetin (n = 12; p < 0.05). This effect was due to the increased NO production, because it was reversed when the cells were treated with a combination of quercetin and l-NMMA. In conclusion quercetin improves endothelial dysfunction by increasing NO synthesis involving BK(Ca)-dependent membrane hyperpolarization-induced capacitative Ca 2+ entry. Increased NO production is responsible for the quercetin-dependent inhibition of endothelial proliferation.
authors.affiliation: Department of Cardiology and Angiology, Justus-Liebig-University of Giessen, Giessen, Germany.
authors.forename: Christoph R
authors.initials: CR
authors.lastname: Kuhlmann
authors.forename: Christian A
authors.initials: CA
authors.lastname: Schaefer
authors.forename: Christoph
authors.initials: C
authors.lastname: Kosok
authors.forename: Yaser
authors.initials: Y
authors.lastname: Abdallah
authors.forename: Sabine
authors.initials: S
authors.lastname: Walther
authors.forename: Dörte W
authors.initials: DW
authors.lastname: Lüdders
authors.forename: Thomas
authors.initials: T
authors.lastname: Neumann
authors.forename: Harald
authors.initials: H
authors.lastname: Tillmanns
authors.forename: Claudia
authors.initials: C
authors.lastname: Schäfer
authors.forename: Hans M
authors.initials: HM
authors.lastname: Piper
authors.forename: Ali
authors.initials: A
authors.lastname: Erdogan
mesh_terms: D002118:Calcium / Q000378:metabolism*; D002455:Cell Division / Q000187:drug effects; D006152:Cyclic GMP / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D006801:Humans; D008564:Membrane Potentials; D009569:Nitric Oxide / Q000096:biosynthesis; D008517:Phytotherapy; D010946:Plants, Medicinal; D024681:Potassium Channels, Calcium-Activated / Q000187:drug effects* / Q000502:physiology; D011794:Quercetin / Q000008:administration & dosage / Q000494:pharmacology* / Q000627:therapeutic use
chemical_list: D024681:Potassium Channels, Calcium-Activated; D009569:Nitric Oxide; D011794:Quercetin; D006152:Cyclic GMP; D002118:Calcium