title: Critical role of neutral cholesteryl ester hydrolase 1 in cholesteryl ester hydrolysis in murine macrophages.
abstract: Hydrolysis of intracellular cholesteryl ester (CE) is the rate-limiting step in the efflux of cholesterol from macrophage foam cells. In mouse peritoneal macrophages (MPMs), this process is thought to involve several enzymes: hormone-sensitive lipase (Lipe), carboxylesterase 3 (Ces3), neutral CE hydrolase 1 (Nceh1). However, there is some disagreement over the relative contributions of these enzymes. To solve this problem, we first compared the abilities of several compounds to inhibit the hydrolysis of CE in cells overexpressing Lipe, Ces3, or Nceh1. Cells overexpressing Ces3 had negligible neutral CE hydrolase activity. We next examined the effects of these inhibitors on the hydrolysis of CE and subsequent cholesterol trafficking in MPMs. CE accumulation was increased by a selective inhibitor of Nceh1, paraoxon, and two nonselective inhibitors of Nceh1, (+)-AS115 and (-)-AS115, but not by two Lipe-selective inhibitors, orlistat and 76-0079. Paraoxon inhibited cholesterol efflux to apoA-I or HDL, while 76-0079 did not. These results suggest that Nceh1 plays a dominant role over Lipe in the hydrolysis of CE and subsequent cholesterol efflux in MPMs.
authors.affiliation: Division of Endocrinology and Metabolism, Department of Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
authors.forename: Kent
authors.initials: K
authors.lastname: Sakai
authors.affiliation: Department of Metabolic Diseases, Graduate School of Medicine, University of Tokyo, Tokyo 113-8655 Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Igarashi
authors.affiliation: Division of Endocrinology and Metabolism, Department of Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Yamamuro
authors.affiliation: Division of Endocrinology and Metabolism, Department of Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
authors.forename: Taichi
authors.initials: T
authors.lastname: Ohshiro
authors.affiliation: Division of Endocrinology and Metabolism, Department of Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
authors.forename: Shuichi
authors.initials: S
authors.lastname: Nagashima
authors.affiliation: Division of Endocrinology and Metabolism, Department of Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Takahashi
authors.affiliation: Division of Endocrinology and Metabolism, Department of Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
authors.forename: Bolormaa
authors.initials: B
authors.lastname: Enkhtuvshin
authors.affiliation: Department of Metabolic Diseases, Graduate School of Medicine, University of Tokyo, Tokyo 113-8655 Japan.
authors.forename: Motohiro
authors.initials: M
authors.lastname: Sekiya
authors.affiliation: Department of Metabolic Diseases, Graduate School of Medicine, University of Tokyo, Tokyo 113-8655 Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Okazaki
authors.affiliation: Division of Endocrinology and Metabolism, Department of Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Osuga
authors.affiliation: Division of Endocrinology and Metabolism, Department of Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
authors.forename: Shun
authors.initials: S
authors.lastname: Ishibashi
mesh_terms: D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects / Q000235:genetics; D043182:Carboxylesterase / Q000235:genetics / Q000378:metabolism; D002788:Cholesterol Esters / Q000235:genetics / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D057809:HEK293 Cells; D006801:Humans; D006868:Hydrolysis; D017737:Macrophages, Peritoneal / Q000201:enzymology*; D051379:Mice; D018345:Mice, Knockout; D002787:Sterol Esterase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D002788:Cholesterol Esters; D004791:Enzyme Inhibitors; C514712:Nceh1 protein, mouse; D043182:Carboxylesterase; D002787:Sterol Esterase
keywords: ATP binding cassette transporters; efflux; foam cells; hormone-sensitive lipase; inhibitor; lipoproteins; paraoxon