Effects of a common human gene variant of extracellular superoxide dismutase on endothelial function after endotoxin in mice.
A common gene variant in the heparin-binding domain (HBD) of extracellular superoxide dismutase (ECSOD) may predispose human carriers to ischaemic heart disease. We have demonstrated that the HBD of ECSOD is important for ECSOD to restore vascular dysfunction produced by endotoxin. The purpose of this study was to determine whether the gene variant in the HBD of ECSOD (ECSOD(R213G)) protects against endothelial dysfunction in a model of inflammation. We constructed a recombinant adenovirus that expresses ECSOD(R213G). Adenoviral vectors expressing ECSOD, ECSOD(R213G) or beta-galactosidase (LacZ, a control) were injected i.v. in mice. After 3 days, at which time the plasma SOD activity is maximal, vehicle or endotoxin (lipopolysaccharide or LPS, 40 mg kg(-1)) was injected i.p. Vasomotor function of aorta in vitro was examined 1 day later. Maximal relaxation to sodium nitroprusside was similar in aorta from normal and LPS-treated mice. Maximal relaxation to acetylcholine (10(-5)) was impaired after LPS and LacZ (63 +/- 3%, mean +/- s.e.m.) compared to normal vessels (83 +/- 3%) (P < 0.05). Gene transfer of ECSOD improved (P < 0.05) relaxation in response to acetylcholine (76 +/- 5%) after LPS, whereas gene transfer of ECSOD(R213G) had no effect (65 +/- 4%). Superoxide was increased in aorta (measured using lucigenin and hydroethidine) after LPS, and levels of superoxide were significantly reduced following ECSOD but not ECSOD(R213G). Thus, ECSOD reduces superoxide and improves relaxation to acetylcholine in the aorta after LPS, while the ECSOD variant R213G had minimal effect. These findings suggest that, in contrast to ECSOD, the common human gene variant of ECSOD fails to protect against endothelial dysfunction produced by an inflammatory stimulus.
Lund
Donald D
DD
Department of Internal Medicine, Cardiovascular Center, University of Iowa Carver College of Medicine, Iowa City, Iowa 52242, USA.
Chu
Yi
Y
Brooks
Robert M
RM
Faraci
Frank M
FM
Heistad
Donald D
DD
D000109:Acetylcholine / Q000494:pharmacology; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001011:Aorta / Q000201:enzymology / Q000503:physiopathology; D015237:Dinoprost / Q000494:pharmacology; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000187:drug effects / Q000201:enzymology / Q000503:physiopathology*; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D007249:Inflammation / Q000201:enzymology / Q000503:physiopathology*; D007763:Lac Operon; D008070:Lipopolysaccharides; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009599:Nitroprusside / Q000494:pharmacology; D011110:Polymorphism, Genetic; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism; D014661:Vasoconstriction / Q000187:drug effects; D014662:Vasoconstrictor Agents / Q000494:pharmacology; D014664:Vasodilation / Q000187:drug effects; D014665:Vasodilator Agents / Q000494:pharmacology; D001616:beta-Galactosidase
D008070:Lipopolysaccharides; D014662:Vasoconstrictor Agents; D014665:Vasodilator Agents; D013481:Superoxides; D009599:Nitroprusside; D015237:Dinoprost; C441642:SOD3 protein, human; D013482:Superoxide Dismutase; D001616:beta-Galactosidase; D000109:Acetylcholine
Am J Physiol Heart Circ Physiol. 2000 Oct;279(4):H1555-62
11009441
Circ Res. 2000 Sep 29;87(7):E18-24
11009571
Free Radic Biol Med. 2002 Mar 1;32(5):446-53
11864784
Methods Enzymol. 2002;346:263-76
11883072
Cardiovasc Res. 2002 Aug 1;55(2):239-49
12123763
Circulation. 2002 Nov 5;106(19):2497-502
12417549
Stroke. 2003 Feb;34(2):434-40
12574556
Circ Res. 2003 Mar 7;92(4):461-8
12600899
Shock. 2003 May;19(5):486-93
12744495
Br J Pharmacol. 2003 May;139(2):444-56
12770950
Circ Res. 2003 Oct 3;93(7):622-9
12933702
Circulation. 2004 Jan 6;109(1):59-65
14662715
Stroke. 2004 Aug;35(8):1963-7
15218160
Am J Physiol Heart Circ Physiol. 2004 Aug;287(2):H805-11
15277203
Mol Ther. 2004 Aug;10(2):344-54
15294181
Clin Chim Acta. 1982 Nov 24;126(1):41-51
7172448
Proc Natl Acad Sci U S A. 1982 Dec;79(24):7634-8
6961438
Eur J Pharmacol. 1990 Nov 6;190(1-2):185-92
1706272
Hum Mol Genet. 1994 Dec;3(12):2251-4
7881430
Biochem J. 1996 Jan 1;313 ( Pt 1):235-9
8546689
J Intern Med. 1997 Jul;242(1):5-14
9260561
J Physiol Pharmacol. 1997 Dec;48(4):633-44
9444612
Br J Pharmacol. 1998 Feb;123(3):525-37
9504394
Circ Res. 1998 Jun 29;82(12):1298-305
9648726
Am J Physiol. 1998 Aug;275(2 Pt 2):H416-21
9683428
Stroke. 1999 Oct;30(10):2191-5; discussion 2195-6
10512928
Am J Physiol Heart Circ Physiol. 2005 Aug;289(2):H624-30
16014616
Mol Ther. 2005 Aug;12(2):321-7
16043100
Circulation. 2005 Aug 16;112(7):1047-53
16087794
Am J Physiol Heart Circ Physiol. 2006 Aug;291(2):H914-20
16840738
Hypertension. 2000 Feb;35(2):595-601
10679503
Nephron. 2000 Mar;84(3):218-23
10720891
Pharmacol Res. 2001 Jul;44(1):22-7
11428906