Abstract
Although active oxygen species play important roles in the pathogenesis of various diseases, the molecular mechanism for oxygen toxicity in vascular diseases remains to be elucidated. Since endothelium-derived relaxing factor (EDRF) is inactivated by superoxide radicals in vitro, oxidative stress in and around vascular endothelial cells may affect the circulatory status of animals. To study the role of superoxide radicals and related enzymes, such as superoxide dismutase (SOD), in vascular diseases, we have developed a fusion protein (HB-SOD) consisting of human Cu/Zn-type SOD and a C-terminal basic peptide with high affinity for heparan sulfate on endothelial cells. When injected intravenously, HB-SOD bound to vascular endothelial cells, underwent transcellular transport, and localized within vascular walls by a heparin-inhibitable mechanism. The blood pressure of spontaneously hypertensive rats (SHR) but not normal animals was decreased significantly by HB-SOD. Heparin inhibited the depressor effect of HB-SOD. In contrast, native SOD had no effect on blood pressure of either SHR or normal rats. Neither H2O2-inactivated HB-SOD nor the C-terminal heparin-binding peptide showed such a depressor effect, suggesting that the catalytic function of HB-SOD is responsible for its depressor action. To know the source of superoxide radicals, we determined xanthine oxidase activity in the aorta and uric acid levels in the plasma. Although no appreciable difference in xanthine oxidase activity was found between the two animal groups, uric acid levels were significantly higher in SHR than in normal rats. Oxypurinol, a potent inhibitor of xanthine oxidase, also decreased the blood pressure of SHR but not of normal rats. These findings indicate that superoxide radicals in and around vascular endothelial cells play critical roles in the pathogenesis of hypertension of SHR.
MeSH Terms
Amino Acid Sequence
Animals
Aorta/drug effects,physiology
Binding Sites
Blood Pressure/drug effects
Heparin/metabolism
Humans
Hypertension/etiology,physiopathology
Immunoenzyme Techniques
Male
Molecular Sequence Data
Muscle, Smooth, Vascular/drug effects,physiology
Oxypurinol/pharmacology
Peptides/chemical synthesis,isolation & purification,metabolism,pharmacology
Rats
Rats, Inbred SHR
Rats, Inbred WKY
Recombinant Proteins/isolation & purification,metabolism,pharmacology
Superoxide Dismutase/genetics,isolation & purification,metabolism,pharmacology
Superoxides/metabolism
Chemicals
Peptides
Recombinant Proteins
Superoxides
Heparin
Superoxide Dismutase
Oxypurinol
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Nakazono K
Department of Biochemistry, Kumamoto University Medical School, Japan.
Watanabe N
Matsuno K
Sasaki J
Sato T
Inoue M
References (26)
26 references, click to expand
-
Development of a strain of spontaneously hypertensive rats.
Jpn Circ J. 1963 Mar;27:282-93
PMID: 13939773
-
Hyperuricemia in primary and renal hypertension.
N Engl J Med. 1966 Sep 1;275(9):457-64
PMID: 5917940
-
Pressor effect of NG-monomethyl-L-arginine in SHRSP.
Jpn J Pharmacol. 1990 Dec;54(4):461-3
PMID: 2087008
-
Expression of a hybrid Cu/Zn-type superoxide dismutase which has high affinity for heparin-like proteoglycans on vascular endothelial cells.
J Biol Chem. 1991 Sep 5;266(25):16409-14
PMID: 1885572
-
Mechanisms of conversion of xanthine dehydrogenase to xanthine oxidase in ischemic rat liver and kidney.
Am J Physiol. 1988 May;254(5 Pt 1):G753-60
PMID: 3163235
-
Identification of arginine as a precursor of endothelium-derived relaxing factor.
Proc Natl Acad Sci U S A. 1988 Nov;85(22):8664-7
PMID: 3263652
-
Superoxide anion is involved in the breakdown of endothelium-derived vascular relaxing factor.
Nature. 1986 Apr 3-9;320(6061):454-6
PMID: 3007998
-
Superoxide anions and hyperoxia inactivate endothelium-derived relaxing factor.
Am J Physiol. 1986 May;250(5 Pt 2):H822-7
PMID: 3010744
-
Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor.
Nature. 1987 Jun 11-17;327(6122):524-6
PMID: 3495737
-
Role of endothelium-derived nitric oxide in the regulation of blood pressure.
Proc Natl Acad Sci U S A. 1989 May;86(9):3375-8
PMID: 2497467
-
Role of superoxide in endothelial-cell modification of low-density lipoproteins.
Biochim Biophys Acta. 1988 Mar 4;959(1):20-30
PMID: 2830901
-
Measurement of endothelial cell free radical generation: evidence for a central mechanism of free radical injury in postischemic tissues.
Proc Natl Acad Sci U S A. 1988 Jun;85(11):4046-50
PMID: 2836868
-
The primary localization of free radical generation after anoxia/reoxygenation in isolated endothelial cells.
Surgery. 1987 Aug;102(2):122-31
PMID: 3039675
-
Superoxide dismutase and catalase conjugated to polyethylene glycol increases endothelial enzyme activity and oxidant resistance.
J Biol Chem. 1988 May 15;263(14):6884-92
PMID: 3129432
-
Basic polyamino acids rich in arginine, lysine, or ornithine cause both enhancement of and refractoriness to formation of endothelium-derived nitric oxide in pulmonary artery and vein.
Circ Res. 1989 Feb;64(2):315-29
PMID: 2492213
-
Splenic outer periarterial lymphoid sheath (PALS): an immunoproliferative microenvironment constituted by antigen-laden marginal metallophils and ED2-positive macrophages in the rat.
Cell Tissue Res. 1989 Sep;257(3):459-70
PMID: 2790931
-
Synthesis of a superoxide dismutase derivative that circulates bound to albumin and accumulates in tissues whose pH is decreased.
Biochemistry. 1989 Aug 8;28(16):6619-24
PMID: 2790016
-
L-arginine is the physiological precursor for the formation of nitric oxide in endothelium-dependent relaxation.
Biochem Biophys Res Commun. 1988 Jun 30;153(3):1251-6
PMID: 3390182
-
Significance of xanthine oxidase in capillary endothelial cells.
Acta Physiol Scand Suppl. 1986;548:39-46
PMID: 3463124
-
Inhibition of oxygen toxicity by targeting superoxide dismutase to endothelial cell surface.
FEBS Lett. 1990 Aug 20;269(1):89-92
PMID: 2387419
-
Immunocytochemical localization of gamma-glutamyltransferase in rat kidney with protein A-horseradish peroxidase.
Proc Natl Acad Sci U S A. 1982 Jun;79(11):3547-50
PMID: 6124972
-
Localization of xanthine oxidase in mammary-gland epithelium and capillary endothelium.
Cell. 1981 Jul;25(1):67-82
PMID: 6895049
-
Enzyme electrodes and solid surface fluorescence methods.
Methods Enzymol. 1976;44:579-633
PMID: 15191
-
Hypertension and hyperuricaemia.
Lancet. 1966 Jan 1;1(7427):15-8
PMID: 4159153
-
Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein).
J Biol Chem. 1969 Nov 25;244(22):6049-55
PMID: 5389100
-
Protein measurement with the Folin phenol reagent.
J Biol Chem. 1951 Nov;193(1):265-75
PMID: 14907713