title: Angiotensin II impairs endothelial nitric-oxide synthase bioavailability under free cholesterol-enriched conditions via intracellular free cholesterol-rich membrane microdomains.
abstract: Vascular endothelial function is impaired in hypercholesterolemia partly because of injury by modified LDL. In addition to modified LDL, free cholesterol (FC) is thought to play an important role in the development of endothelial dysfunction, although the precise mechanisms remain to be elucidated. The aim of this study was to clarify the mechanisms of endothelial dysfunction induced by an FC-rich environment. Loading cultured human aortic endothelial cells with FC induced the formation of vesicular structures composed of FC-rich membranes. Raft proteins such as phospho-caveolin-1 (Tyr-14) and small GTPase Rac were accumulated toward FC-rich membranes around vesicular structures. In the presence of these vesicles, angiotensin II-induced production of reactive oxygen species (ROS) was considerably enhanced. This ROS shifted endothelial NOS (eNOS) toward vesicle membranes and vesicles with a FC-rich domain trafficked toward perinuclear late endosomes/lysosomes, which resulted in the deterioration of eNOS Ser-1177 phosphorylation and NO production. Angiotensin II-induced ROS decreased the bioavailability of eNOS under the FC-enriched condition.
authors.affiliation: Department of Cardiovascular Medicine, Graduate School of Medicine, the University of Tokyo, Tokyo 113-8655, Japan.
authors.forename: Eisuke
authors.initials: E
authors.lastname: Amiya
authors.affiliation: Department of Cardiovascular Medicine, Graduate School of Medicine, the University of Tokyo, Tokyo 113-8655, Japan.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Watanabe
authors.affiliation: Department of Cardiovascular Medicine, Graduate School of Medicine, the University of Tokyo, Tokyo 113-8655, Japan.
authors.forename: Norihiko
authors.initials: N
authors.lastname: Takeda
authors.affiliation: Department of Cardiovascular Medicine, Graduate School of Medicine, the University of Tokyo, Tokyo 113-8655, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Saito
authors.affiliation: Department of Cardiovascular Medicine, Graduate School of Medicine, the University of Tokyo, Tokyo 113-8655, Japan.
authors.forename: Taro
authors.initials: T
authors.lastname: Shiga
authors.affiliation: Department of Cardiovascular Medicine, Graduate School of Medicine, the University of Tokyo, Tokyo 113-8655, Japan.
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Hosoya
authors.affiliation: Department of Cardiovascular Medicine, Graduate School of Medicine, the University of Tokyo, Tokyo 113-8655, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Nakao
authors.affiliation: Department of Cardiovascular Medicine, Graduate School of Medicine, the University of Tokyo, Tokyo 113-8655, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Imai
authors.affiliation: Department of Cardiovascular Medicine, Graduate School of Medicine, the University of Tokyo, Tokyo 113-8655, Japan.
authors.forename: Ichiro
authors.initials: I
authors.lastname: Manabe
authors.affiliation: Department of Cardiovascular Medicine, Graduate School of Medicine, the University of Tokyo, Tokyo 113-8655, Japan; Jichi Medical University, Shimotsuke 329-0498, Japan.
authors.forename: Ryozo
authors.initials: R
authors.lastname: Nagai
authors.affiliation: Department of Cardiovascular Medicine, Graduate School of Medicine, the University of Tokyo, Tokyo 113-8655, Japan; Department of Cardiovascular Medicine, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan.
authors.forename: Issei
authors.initials: I
authors.lastname: Komuro
authors.affiliation: Department of Cardiovascular Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki 852-8102, Japan. Electronic address: maemura@nagasaki-u.ac.jp.
authors.forename: Koji
authors.initials: K
authors.lastname: Maemura
mesh_terms: D000804:Angiotensin II / Q000378:metabolism*; D001011:Aorta / Q000166:cytology; D001682:Biological Availability; D002784:Cholesterol / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism; D006801:Humans; D008247:Lysosomes / Q000378:metabolism; D021962:Membrane Microdomains / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D017382:Reactive Oxygen Species / Q000378:metabolism; D012694:Serine / Q000378:metabolism
chemical_list: D017382:Reactive Oxygen Species; D000804:Angiotensin II; D009569:Nitric Oxide; D012694:Serine; D002784:Cholesterol; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III
keywords: Angiotensin II; Cholesterol; Endothelial Dysfunction; Nitric-oxide Synthase; Reactive Oxygen Species (ROS)
references.citation: Circulation. 2004 Jun 1;109(21 Suppl 1):II27-33
references.pmid: 15173060
references.citation: J Clin Invest. 2008 Nov;118(11):3701-13
references.pmid: 18924609
references.citation: Am J Pathol. 1986 Apr;123(1):109-25
references.pmid: 3963146
references.citation: Am J Pathol. 1988 Apr;131(1):73-83
references.pmid: 3354646
references.citation: FASEB J. 2006 Jul;20(9):1501-3
references.pmid: 16754746
references.citation: J Cell Biol. 1983 May;96(5):1273-8
references.pmid: 6132922
references.citation: J Neurosci. 2004 Apr 21;24(16):4070-81
references.pmid: 15102922
references.citation: J Inherit Metab Dis. 2007 Feb;30(1):79-87
references.pmid: 17189993
references.citation: Biochim Biophys Acta. 2007 Jun;1768(6):1311-24
references.pmid: 17493580
references.citation: Arterioscler Thromb Vasc Biol. 2003 Aug 1;23(8):1430-6
references.pmid: 12750117
references.citation: Circ Res. 2006 Aug 4;99(3):299-306
references.pmid: 16825579
references.citation: Circulation. 2005 Feb 8;111(5):629-32
references.pmid: 15668341
references.citation: EMBO J. 2002 Aug 1;21(15):3989-4000
references.pmid: 12145200
references.citation: Vascul Pharmacol. 2002 May;38(5):275-82
references.pmid: 12487032
references.citation: J Lipid Res. 2000 Feb;41(2):226-37
references.pmid: 10681406
references.citation: J Biol Chem. 2001 May 25;276(21):18507-12
references.pmid: 11279173
references.citation: Arterioscler Thromb Vasc Biol. 2009 Oct;29(10):1615-21
references.pmid: 19556523
references.citation: Ann Med. 2005;37(6):394-403
references.pmid: 16203612
references.citation: Arterioscler Thromb Vasc Biol. 2011 Sep;31(9):2098-105
references.pmid: 21659644
references.citation: Proc Natl Acad Sci U S A. 2009 Oct 13;106(41):17343-8
references.pmid: 19805165
references.citation: Arterioscler Thromb Vasc Biol. 2006 Feb;26(2):372-8
references.pmid: 16306428
references.citation: Proc Natl Acad Sci U S A. 2003 Nov 11;100(23):13531-6
references.pmid: 14581613
references.citation: J Lipid Res. 1999 Sep;40(9):1728-33
references.pmid: 10484621
references.citation: J Cell Biol. 1995 Dec;131(6 Pt 1):1421-33
references.pmid: 8522601
references.citation: J Neurosci. 2012 Apr 18;32(16):5573-84
references.pmid: 22514319
references.citation: Free Radic Biol Med. 2002 Jul 1;33(1):52-62
references.pmid: 12086682
references.citation: J Lipid Res. 1991 Jul;32(7):1125-31
references.pmid: 1940636
references.citation: Am J Physiol Cell Physiol. 2008 Nov;295(5):C1281-91
references.pmid: 18787079
references.citation: Circ Res. 2002 May 3;90(8):866-73
references.pmid: 11988487
references.citation: Biochemistry. 2010 Apr 13;49(14):3129-37
references.pmid: 20184376
references.citation: Proc Natl Acad Sci U S A. 2004 Feb 24;101(8):2619-24
references.pmid: 14983058
references.citation: Circ J. 2012;76(11):2497-512
references.pmid: 23075817
references.citation: Nat Cell Biol. 2003 Sep;5(9):781-92
references.pmid: 12907943
references.citation: FASEB J. 2006 Dec;20(14):2565-6
references.pmid: 17060403
references.citation: J Clin Invest. 2001 Nov;108(9):1341-8
references.pmid: 11696579
references.citation: Mol Genet Metab. 2010 Feb;99(2):99-108
references.pmid: 19900828
references.citation: Histochem Cell Biol. 2001 Jul;116(1):63-8
references.pmid: 11479724
references.citation: Proc Natl Acad Sci U S A. 2009 Apr 21;106(16):6849-53
references.pmid: 19342481
references.citation: Arterioscler Thromb Vasc Biol. 2008 Mar;28(3):485-90
references.pmid: 18162606
references.citation: Histochem Cell Biol. 2001 Aug;116(2):119-30
references.pmid: 11685540
references.citation: J Biol Chem. 1999 Nov 5;274(45):32512-9
references.pmid: 10542298
references.citation: Circ Res. 2007 Jun 8;100(11):1634-42
references.pmid: 17478727
references.citation: Pharmacol Rev. 1996 Mar;48(1):3-19
references.pmid: 8685247
references.citation: Am J Physiol Cell Physiol. 2011 Apr;300(4):C927-36
references.pmid: 21228321
references.citation: Circ Res. 2006 Apr 28;98(8):1064-71
references.pmid: 16556870
references.citation: Free Radic Res. 2010 Aug;44(8):831-42
references.pmid: 20528560
references.citation: Arterioscler Thromb Vasc Biol. 2005 Sep;25(9):1824-30
references.pmid: 15976327
references.citation: Atherosclerosis. 1984 May-Jun;51(2-3):281-92
references.pmid: 6204652