title: Cytochrome P-450 metabolites but not NO, PGI2, and H2O2 contribute to ACh-induced hyperpolarization of pressurized canine coronary microvessels.
abstract: The endothelium-dependent hyperpolarization of cells has a crucial role in regulating vascular tone, especially in microvessels. Nitric oxide (NO) and prostacyclin (PGI2), in addition to endothelium-derived hyperpolarizing factor (EDHF), have been reported to hyperpolarize vascular smooth muscle in several organs. Studies have reported the hyperpolarizing effects of these factors are increased by a stretch in large coronary arteries. EDHF has not yet been identified and cytochrome P-450 metabolites and H2O2 are candidates for EDHF. With the use of the membrane potential-sensitive fluorescent dye bis-(1,3-dibutylbarbituric acid)trimethione oxonol [DiBAC4(3)], we examined whether NO, PGI2, cytochrome P-450 metabolites, and H2O2 contribute to ACh-induced hyperpolarization in pressurized coronary microvessels. Canine coronary arterial microvessels (60-356 mum internal diameter) were cannulated and pressurized at 60 cmH2O in a vessel chamber perfused with physiological salt solution containing DiBAC4(3). Fluorescence intensity and diameter were measured on a computer. There was a linear correlation between changes in the fluorescence intensity and membrane potential. ACh significantly decreased the fluorescence intensity (hyperpolarization) of the microvessels without any inhibitors. Endothelial damage caused by air perfusion abolished the ACh-induced decrease in fluorescence intensity. The inhibitors of NO synthase and cyclooxygenase did not affect the ACh-induced decreases in the fluorescence intensity. The addition of 17-octadecynoic acid, a cytochrome P-450 monooxygenase inhibitor, to those inhibitors significantly attenuated the ACh-induced decreases in fluorescence intensity, whereas catalase, an enzyme that dismutates H2O2 to form water and oxygen, did not. Furthermore, catalase did not affect the vasodilation produced by ACh. These results indicate that NO and PGI2 do not contribute to the ACh-induced hyperpolarization and that the cytochrome P-450 metabolites but not H2O2 are involved in EDHF-mediated hyperpolarization in canine coronary arterial microvessels.
authors.affiliation: Department of Comprehensive Medicine, Tohoku University School of Medicine, 1-1 Seiryo-machi, Aobaku, Sendai 980-8574, Japan.
authors.forename: Mitsuaki
authors.initials: M
authors.lastname: Tanaka
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kanatsuka
authors.forename: Boon-Hooi
authors.initials: BH
authors.lastname: Ong
authors.forename: Toshinori
authors.initials: T
authors.lastname: Tanikawa
authors.forename: Akira
authors.initials: A
authors.lastname: Uruno
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Komaru
authors.forename: Ryoji
authors.initials: R
authors.lastname: Koshida
authors.forename: Kunio
authors.initials: K
authors.lastname: Shirato
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D003326:Coronary Circulation / Q000187:drug effects / Q000502:physiology*; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004285:Dogs; D004730:Endothelium, Vascular / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D011464:Epoprostenol / Q000378:metabolism*; D005260:Female; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D008833:Microcirculation / Q000187:drug effects / Q000502:physiology; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D016877:Oxidants / Q000494:pharmacology*; D011188:Potassium / Q000494:pharmacology; D011312:Pressure; D014665:Vasodilator Agents / Q000494:pharmacology
chemical_list: D016861:Cyclooxygenase Inhibitors; D065607:Cytochrome P-450 Enzyme Inhibitors; D004791:Enzyme Inhibitors; D016877:Oxidants; D014665:Vasodilator Agents; D009569:Nitric Oxide; D003577:Cytochrome P-450 Enzyme System; D006861:Hydrogen Peroxide; D011464:Epoprostenol; D019001:Nitric Oxide Synthase; D000109:Acetylcholine; D011188:Potassium