title: Nitric oxide derived from human umbilical vein endothelial cells inhibits transendothelial migration of neutrophils.
abstract: We evaluated the roles of nitric oxide (NO) derived from endothelial cells in neutrophil transendothelial migration (TEM). Pretreatment of human umbilical vein endothelial cells (HUVECs) with NG-nitro-L-arginine methyl ester hydrochloride (L-NAME) or NG-monomethyl L-arginine (L-NMMA), which are inhibitors of NO synthases, enhanced neutrophil TEM. Similar augmentation of TEM was observed in the presence of an NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy PTIO). Neutrophil TEM across L-NAME- or L-NMMA-treated HUVECs was inhibited by continuous NO supply by NO donors. These findings support the suggestion that continuous production of NO by endothelial cells suppresses neutrophil TEM. Flow cytometric analyses revealed that NO accumulates in neutrophils co-cultured with NO-producing HUVECs. A decreased amount of NO was detected in neutrophils co-cultured with L-NAME-treated HUVECs compared with neutrophils co-cultured with untreated HUVECs. Soluble guanylyl cyclase (sGC) is known as one of the most important targets of NO in neutrophils. 3-(53-Hydroxymethyl-23furyl)-1-benzyl indazole (YC-1), an activator of sGC, inhibited L-NAME-induced neutrophil TEM. It was interesting that inhibition of neutrophil sGC with 1-H[1,2,4-]oxadiazolo[4,3-a]quinoxalin-1-1 (ODQ) was sufficient to enhance TEM. These results suggest that NO derived from HUVECs acts on neutrophils to inhibit TEM, at least in part by activating sGC. Our findings imply the role of NO constitutively generated by HUVECs in protection against excessive neutrophil extravasation and unnecessary tissue damage under physiological conditions.
authors.affiliation: Department of Hematology and Oncology, Graduate School of Medicine, Kyoto University, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Oka
authors.forename: Masataka
authors.initials: M
authors.lastname: Sasada
authors.forename: Kokichi
authors.initials: K
authors.lastname: Yamamoto
authors.forename: Masaharu
authors.initials: M
authors.lastname: Nohgawa
authors.forename: Atsushi
authors.initials: A
authors.lastname: Takahashi
authors.forename: Kouhei
authors.initials: K
authors.lastname: Yamashita
authors.forename: Hiroko
authors.initials: H
authors.lastname: Yamada
authors.forename: Takashi
authors.initials: T
authors.lastname: Uchiyama
mesh_terms: D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D009504:Neutrophils / Q000502:physiology*; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D014471:Umbilical Veins / Q000166:cytology / Q000502:physiology*
chemical_list: D004791:Enzyme Inhibitors; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase