Abstract
The superoxide anion (O2·−) appears to be an important modulator of nitric oxide bioavailability. Enzymatic scavenging of O2·− is carried out by superoxide dismutase (SOD). The present study was designed to characterize the developmental changes on pulmonary vascular reactivity induced by 1) exogenous Cu/Zn SOD, 2) several putative SOD mimetics, and 3) endogenous SOD inhibition. We also analyzed age-related changes on pulmonary SOD activity and vascular O2·− levels. SOD (1–300 U/mL) produced endothelium-dependent relaxation of U46619-contracted intrapulmonary arteries (fourth branch) and veins from 12- to 24-h-old and 2-wk-old piglets. SOD-induced relaxation was greater in pulmonary arteries and was abolished by the nitric oxide synthase inhibitor Nω-nitro-l-arginine methyl ester. SOD induced a greater pulmonary artery relaxation in the 2-wk-old than in the 12- to 24-h-old piglet. SOD (100 U/mL) did not modify acetylcholine-induced relaxation in pulmonary arteries. In contrast, endogenous SOD inhibition by diethyldithiocarbamate (3 mM) impaired acetylcholine-induced relaxation in pulmonary arteries from newborn but not from 2-wk-old piglets. Total SOD activity in lung tissue did not change with postnatal age. With the use of dihydroethidium, an oxidant-sensitive fluorescent probe, we did not find significant age- or vessel-related differences in O2·− presence. From the putative SOD mimetics tested, only the metal salts MnCl2 and CuSO4 reproduced the vascular effects of SOD. In summary, SOD produces endothelium-dependent pulmonary vascular relaxation by protecting nitric oxide from destruction by O2·−. This effect was less marked in newborns than in 2-wk-old piglets. In contrast, pulmonary arteries from newborn piglets are more sensitive to the inhibition of endogenous SOD.
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Abbreviations
- ACh:
-
acetylcholine
- DETCA:
-
diethyldithiocarbamate
- DHE:
-
dihydroethidium
- l-NAME:
-
Nω-nitro-l-arginine methyl ester
- MnTMPyP:
-
Mn [III] tetrakis [1-methyl-4-pyridyl] porphyrin
- NBT:
-
nitro blue tetrazolium
- NO·:
-
nitric oxide
- ONOO−:
-
peroxynitrite
- PEG-SOD:
-
polyethylene glycosylated SOD
- PPHN:
-
persistent pulmonary hypertension of the newborn
- PTIYO:
-
4-phenyl-2,2,5,5-tetramethyl imidazolin-1-yloxy-5-oxide
- SOD:
-
superoxide dismutase
- U46619:
-
9,11-dideoxy-11α,9α-epoxymethano-prostaglandin F2α
References
Abman SH, Stevens T 1996 Perinatal pulmonary vasoregulation: implications for the pathophysiology and treatment of neonatal pulmonary hypertension. In: Haddad G, Lister G (eds) Tissue Oxygen Deprivation: Developmental, Molecular and Integrative Function. Marcel Dekker, New York, pp 367–432.
Beckman JS, Koppenol WH 1996 Nitric oxide, superoxide, and peroxynitrite: the good, the bad, and the ugly. Am J Physiol 271: C1424–C1437
Sanderud J, Norstein J, Saugstad OD 1991 Reactive oxygen metabolites produce pulmonary vasoconstriction in young pigs. Pediatr Res 29: 543–547
Liu Q, Wiener CM, Flavahan NA 1998 Superoxide and endothelium-dependent constriction to flow in porcine small pulmonary arteries. Br J Pharmacol 124: 331–336
Walther FJ, Wade AB, Warburton D, Forman HJ 1991 Ontogeny of antioxidant enzymes in the fetal lamb lung. Exp Lung Res 17: 39–45
Frank L 1998 Development of the antioxidant defenses in fetal life. Semin Neonatol 3: 173–182
Asikainen TM, Raivio KO, Saksela M, Kinnula VL 1988 Expression and developmental profile of antioxidant enzymes in human lung and liver. Am J Respir Cell Mol Biol 19: 942–949
Morecroft I, MacLean MR 1998 Developmental changes in endothelium-dependent vasodilation and the influence of superoxide anions in perinatal rabbit pulmonary arteries. Br J Pharmacol 125: 1585–1593
Steinhorn RH, Albert G, Swartz DD, Russell JA, Levine CR, Davis JM 2001 Recombinant human superoxide dismutase enhances the effect of inhaled nitric oxide in persistent pulmonary hypertension. Am J Respir Crit Care Med 164: 834–839
Carpenter D, Larkin H, Chang A, Morris E, O'Neill J, Curtis J 2001 Superoxide dismutase and catalase do not affect the pulmonary hypertensive response to group B streptococcus in the lamb. Pediatr Res 49: 181–188
Villamor E, Perez-Vizcaino F, Cogolludo AL, Conde-Oviedo J, Zaragoza-Arnaez F, Lopez-Lopez JG, Tamargo J 2000 Relaxant effects of carbon monoxide compared with nitric oxide in pulmonary and systemic vessels of newborn piglets. Pediatr Res 48: 546–553
Mian KB, Martin W 1995 Differential sensitivity of basal and acetylcholine-stimulated activity of nitric oxide to destruction by superoxide anion in rat aorta. Br J Pharmacol 115: 993–1000
MacKenzie A, Filippini S, Martin W 1999 Effects of superoxide dismutase mimetics on the activity of nitric oxide in rat aorta. Br J Pharmacol 127: 1159–1164
Villamor E, PerezVizcaino F, Tamargo J, Moro M 1996 Effects of group B streptococcus on the responses to U46619, endothelin-1, and noradrenaline in isolated pulmonary and mesenteric arteries of piglets. Pediatr Res 40: 827–833
MacKenzie A, Martin W 1998 Loss of endothelium-derived nitric oxide in rabbit aorta by oxidant stress: restoration by superoxide dismutase mimetics. Br J Pharmacol 124: 719–728
Beauchamp C, Fridovich I 1971 Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44: 276–287
Li N, Yi FX, Spurrier JL, Bobrowitz CA, Zou AP 2002 Production of superoxide through NADH oxidase in thick ascending limb of Henle's loop in rat kidney. Am J Physiol Renal Physiol 282: F1111–F1119
Stepp DW, Ou J, Ackerman AW, Welak S, Klick D, Pritchard KA 2002 Native LDL, and minimally oxidized LDL differentially regulate superoxide anion in vascular endothelium in situ. Am J Physiol Heart Circ Physiol 283: H750–H759
Levy M, Tulloh RM, Komai H, Stuart-Smith K, Haworth SG 1995 Maturation of the contractile response and its endothelial modulation in newborn porcine intrapulmonary arteries. Pediatr Res 38: 25–29
Liu SF, Hislop AA, Haworth SG, Barnes PJ 1992 Developmental changes in endothelium-dependent pulmonary vasodilatation in pigs. Br J Pharmacol 106: 324–330
Steinhorn RH, Morin FC, Gugino SF, Giese EC, Russell JA 1993 Developmental differences in endothelium-dependent responses in isolated ovine pulmonary arteries and veins. Am J Physiol 264: H2162–H167
Arrigoni FI, Hislop AA, Pollock JS, Haworth SG, Mitchell JA 2002 Birth upregulates nitric oxide synthase activity in the porcine lung. Life Sci 70: 1609–1620
Nozik-Grayck E, Dieterle CS, Piantadosi CA, Enghild JJ, Oury TD 2000 Secretion of extracellular superoxide dismutase in neonatal lungs. Am J Physiol Lung Cell Mol Physiol 279: L977–L984
Perez-Vizcaino F, Lopez-Lopez JG, Santiago R, Cogolludo A, Zaragoza-Arnaez F, Moreno L, Alonso MJ, Salaices M, Tamargo J 2002 Postnatal maturation in nitric oxide-induced pulmonary artery relaxation involving cyclooxygenase-1 activity. Am J Physiol Lung Cell Mol Physiol 283: L839–L848
Steinhorn RH, Russell JA, Lakshminrusimha S, Gugino SF, Black SM, Fineman JR 2001 Altered endothelium-dependent relaxations in lambs with high pulmonary blood flow and pulmonary hypertension. Am J Physiol Heart Circ Physiol 280: H311–H317
Ignarro LJ, Byrns RE, Buga GM, Wood KS 1987 Endothelium-derived relaxing factor from pulmonary artery and vein possesses pharmacologic and chemical properties identical to those of nitric oxide radical. Circ Res 61: 866–879
Cherry PD, Omar HA, Farrell KA, Stuart JS, Wolin MS 1990 Superoxide anion inhibits cGMP-associated bovine pulmonary arterial relaxation. Am J Physiol 259: H1056–H1062
Mugge A, Elwell JH, Peterson TE, Harrison DG 1991 Release of intact endothelium-derived relaxing factor depends on endothelial superoxide dismutase activity. Am J Physiol 260: C219–C225
Beckman JS, Minor RL, White CW, Repine JE, Rosen GM, Freeman BA 1988 Superoxide dismutase and catalase conjugated to polyethylene glycol increases endothelial enzyme activity and oxidant resistance. J Biol Chem 15: 6884–6892
Lopez-Lopez JG, Perez-Vizcaino F, Cogolludo AL, Ibarra M, Zaragoza-Arnaez F, Tamargo J 2001 Nitric oxide- and nitric oxide donors-induced relaxation and its modulation by oxidative stress in piglet pulmonary arteries. Br J Pharmacol 133: 615–624
Laight DW, Kaw AV, Carrier MJ, Anggard EE 1998 Interaction between superoxide anion and nitric oxide in the regulation of vascular endothelial function. Br J Pharmacol 124: 238–244
Pagano PJ, Clark JK, Cifuentes-Pagano ME, Clark SM, Callis GM, Quinn MT 1997 Localization of a constitutively active, phagocyte-like NADPH oxidase in rabbit aortic adventitia: enhancement by angiotensin II. Proc Natl Acad Sci USA 94: 14483–14488
Das M, Stenmark KR, Dempsey EC 1995 Enhanced growth of fetal and neonatal pulmonary artery adventitial fibroblasts is dependent on protein kinase C. Am J Physiol 269: L660–L667
Kelly DA, Hislop AA, Hall SM, Haworth SG 2002 Correlation of pulmonary arterial smooth muscle structure and reactivity during adaptation to extrauterine life. J Vasc Res 39: 30–40
Allen K, Haworth SG 1988 Human postnatal pulmonary arterial remodeling: ultrastructural studies of smooth muscle cell and connective tissue maturation. Lab Invest 59: 702–709
Raj JU, Hillyard R, Kaapa P, Gropper M, Anderson J 1990 Pulmonary arterial and venous constriction during hypoxia in 3- to 5-wk-old and adult ferrets. J Appl Physiol 69: 2183–2189
Gao Y, Tolsa JF, Botello M, Raj JU 1998 Developmental change in isoproterenol-mediated relaxation of pulmonary veins of fetal and newborn lambs. J Appl Physiol 84: 1535–1539
Schmalfuss CM, Chen LY, Bott JN, Staples ED, Mehta JL 1999 Superoxide anion generation, superoxide dismutase activity, and nitric oxide release in human internal mammary artery and saphenous vein segments. J Cardiovasc Pharmacol Ther 4: 249–257
Grishko V, Solomon M, Wilson GL, LeDoux SP, Gillespie MN 2001 Oxygen radical-induced mitochondrial DNA damage and repair in pulmonary vascular endothelial cell phenotypes. Am J Physiol Lung Cell Mol Physiol 280: L1300–L1308
Finer NN, Barrington KJ 2002 Nitric oxide for respiratory failure in infants born at or near term (Cochrane Review). In: The Cochrane Library, Issue 2, Oxford: Update Software
Cuzzocrea S, Riley DP, Caputi AP, Salvemini D 2001 Antioxidant therapy: a new pharmacological approach in shock, inflammation, and ischemia/reperfusion injury. Pharmacol Rev 53: 135–159
Mok JS, Paisley K, Martin W 1998 Inhibition of nitrergic neurotransmission in the bovine retractor penis muscle by an oxidant stress: effects of superoxide dismutase mimetics. Br J Pharmacol 124: 111–118
Mitchell JB, Samuni A, Krishna MC, DeGraff WG, Ahn MS, Samuni U, Russo A 1990 Biologically active metal-independent superoxide dismutase mimics. Biochemistry 29: 2802–2807
Krishna CM, Liebmann JE, Kaufman D, DeGraff W, Hahn SM, McMurry T, Mitchell JB, Russo A 1992 The catecholic metal sequestering agent 1,2-dihydroxybenzene-3,5-disulfonate confers protection against oxidative cell damage. Arch Biochem Biophys 294: 98–106
Akaike T, Yoshida M, Miyamoto Y, Sato K, Kohno M, Sasamoto K, Miyazaki K, Ueda S, Maeda H 1993 Antagonistic action of imidazolineoxyl N-oxides against endothelium-derived relaxing factor/NO through a radical reaction. Biochemistry 32: 827–832
Gardner PR, Nguyen DD, White CW 1996 Superoxide scavenging by Mn(II/III) tetrakis (1-methyl-4-pyridyl) porphyrin in mammalian cells. Arch Biochem Biophys 325: 20–28
Steinhorn RH, Lakshminrusimha S, Gugino SF, Russell JA 2000 Superoxide dismutase relaxes pulmonary arteries from fetal sheep by a mechanism independent of NO-guanylate cyclase signaling. Nitric Oxide 4: 225–226( abstr)
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The authors thank Dr. Francisco Pérez-Vizcaíno for helpful discussion and valuable comments on the manuscript.
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Villamor, E., Kessels, C., Fischer, M. et al. Role of Superoxide Anion on Basal and Stimulated Nitric Oxide Activity in Neonatal Piglet Pulmonary Vessels. Pediatr Res 54, 372–381 (2003). https://doi.org/10.1203/01.PDR.0000077481.15081.C8
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DOI: https://doi.org/10.1203/01.PDR.0000077481.15081.C8
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