title: Effect of homocysteine on nitric oxide production in coronary microvascular endothelial cells.
abstract: Hyperhomocysteinemia is widely recognized as an independent risk factor for coronary artery vascular disease, although the underlying mechanisms are not well understood. This study aims to investigate the effect of homocysteine on nitric oxide (NO) production in coronary microvascular endothelial cells (CMECs) and putative mechanisms mediating this effect. CMECs were isolated on Langendorff system by collagenase perfusion of hearts from male rats and cultured. The effect of homocysteine (0.01 to 1 mM) on basal and stimulated NO production was evaluated by measuring nitrite in the culture media after incubation with or without N(G)-nitro-L-arginine methyl ester (L-NAME) (1 mM), superoxide dismutase (100 U/mL), or catalase (1000 U/mL) for 24 h. Total nitrite was measured using Griess reaction after reduction of nitrate to nitrite with nitrate reductase. Homocysteine did not affect basal nitrite accumulation; however, it significantly increased the nitrite accumulation induced by the calcium ionophore A23187 or interleukin-1beta only at 1 mM. This effect of homocysteine was significantly inhibited by L-NAME, superoxide dismutase, and catalase. In conclusion, homocysteine increases NO release from stimulated CMECs without affecting basal NO production, which is probably accompanied by increased production of reactive oxygen species. It can be postulated that endothelial cells generate NO in order to minimize the damage caused by homocysteine.
authors.affiliation: Department of Pharmacology and Clinical Pharmacology, School of Medicine, Ege University, Izmir, Turkey. ayse.erol@ege.edu.tr
authors.forename: Ayşe
authors.initials: A
authors.lastname: Erol
authors.forename: Mehtap G
authors.initials: MG
authors.lastname: Cinar
authors.forename: Cenk
authors.initials: C
authors.lastname: Can
authors.forename: Murat
authors.initials: M
authors.lastname: Olukman
authors.forename: Sibel
authors.initials: S
authors.lastname: Ulker
authors.forename: Sezen
authors.initials: S
authors.lastname: Koşay
mesh_terms: D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006710:Homocysteine / Q000494:pharmacology*; D053583:Interleukin-1beta / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000096:biosynthesis*; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D053583:Interleukin-1beta; D007476:Ionophores; D017382:Reactive Oxygen Species; D006710:Homocysteine; D009569:Nitric Oxide; D000001:Calcimycin; D019331:NG-Nitroarginine Methyl Ester