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Ascorbate Prevents the Interaction of Superoxide and Nitric Oxide Only at Very High Physiological Concentrations
Abstract
The bioactivity of nitric oxide (.NO) depends, in part, on its interaction with superoxide. Usually, superoxide dismutase (SOD) preserves .NO bioactivity by limiting the availability of superoxide. Ascorbic acid also effectively scavenges superoxide, but the extent to which this interaction is necessary for intact .NO bioactivity is not known. Therefore, the present study examined the effect of ascorbic acid on .NO bioactivity with isolated rabbit arterial segments. A steady flux of superoxide (1.15 to 2.3 micromol . L-1 . min-1) produced either by pyrogallol autoxidation or a hypoxanthine/xanthine oxidase system inhibited endothelium-derived .NO-mediated arterial relaxation elicited by acetylcholine. This effect of superoxide was completely blocked by SOD (300 IU/mL) and the manganese SOD mimic EUK-8 (300 micromol/L) and partially inhibited by ascorbic acid (10 mmol/L). Lower concentrations of ascorbic acid were ineffective despite scavenging >90% of superoxide. We increased the endogenous flux of superoxide (3.2+/-0.3-fold) by inhibiting vascular copper-zinc SOD with diethyldithiocarbamate. This increased endogenous flux of superoxide produced an impairment of .NO-mediated arterial relaxation that was reversed by EUK-8 (300 micromol/L) but not ascorbic acid (10 mmol/L) despite equivalent scavenging of the endogenous superoxide flux. We used 3-nitrotyrosine formation (from peroxynitrite) as an indicator of .NO interaction with superoxide and found that SOD and EUK-8 compete more effectively with .NO for superoxide than does ascorbic acid. These data indicate that preservation of .NO bioactivity by superoxide scavengers depends not only on superoxide scavenging activity, but also on the rate of superoxide scavenging. Normal extracellular concentrations of ascorbic acid (30 to 150 micromol/L) are not likely to prevent the interaction of .NO with superoxide under physiological conditions.
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“However, in competition with NO, a concentration of vitamin C about 10 mM is needed to block the interaction of NO with the superoxide by effective scavenging of the latter [167].”
From Vitamin C-Sources, Physiological Role, Kinetics, Deficiency, Use, Toxicity, and Determination (2021) · cites this paper as [167]
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“2004a,b) that the dose of ascorbic acid infused in the present study results in supraphysiological plasma concentrations known to scavenge superoxide anions (Frei et al. 1989; Jackson et al. 1998), and acutely reduces systemic markers of oxidative stress including plasma concentrations of oxidized low-density lipoprot…”
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“However, in a therapeutic perspective, it must be remembered that at least some of harmful consequences of free-radical production can be efficiently counteracted by AA only at supraphysiological concentrations [34].”
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Karlıdağ, T., Keleş, E., İlhan, N., Yakclin, S., Kaygusuz, İ., & Yildiz, M. (2005) · Annals of Otology Rhinology & Laryngology · doi:10.1177/000348940511400207 · Cited by 23
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“In this scenario, scavenging of superoxide or its downstream products by low millimolar concentrations of ascorbate [41] could well decrease oxidant-induced increases in endothelial barrier permeability.”
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“Second, ascorbate has to reach high enough levels to counteract the superoxide and NO reactions, occurring at a rate that is 105-fold greater than that at which superoxide reacts with ascorbic acid [19].”
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Ascorbic Acid Infusion Blunts CD40L Upregulation in Patients Undergoing Coronary Stent
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Adjuvant vitamin C for sepsis: mono or triple?
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Serra, P. A., Migheli, R., Rocchitta, G., Taras, M. G., Mura, M. P., Delogu, M. R., Esposito, G., Desole, M. S., MIELE, E., & Miele, M. (2003) · Neuroscience Letters · doi:10.1016/j.neulet.2003.07.022 · Cited by 18
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“Ascorbic acid may either protect NO from destruction by superoxide anion (Dudgeon et al., 1998; Jackson et al., 1998) or scavenge it (Whiteman & Halliwell, 1996).”
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“VC prevents the interaction of O2 and NO at very high physiological concentrations [53] and reacts with ONOO in vitro [54].”
BackgroundVitamin C improves endothelial function in healthy estrogen-deficient postmenopausal women
McSorley, P., Young, I. S., Bell, P. M., Fee, J. P. H., & McCance, D. R. (2003) · Climacteric · doi:10.1080/cmt.6.3.238.247 · Cited by 17
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“Antioxidant treatments effectively reverse the impaired intrahepatic production of NO (Ting et al. 1996; Jackson et al. 1998; Taddei et al. 1998; Hernandez-Guerra et al. 2006; Karaa et al. 2006).”
Protective effects of anthocyanins against brain ischemic damage
Škėmienė, K., Pampuščenko, K., Rekuvienė, E., & Borutaitė, V. (2020) · Journal of Bioenergetics and Biomembranes · doi:10.1007/s10863-020-09825-9 · Cited by 17
“…obtained results with well-known antioxidant – ascorbic acid (AA), in its oxidized form dehydroascorbic acid (DHA), which penetrates the blood–brain barrier through glucose transporters and accumulates within brain cells (Agus et al. 1997; Jackson et al. 1998; Vera et al. 1993; Vera et al. 1995).”
BackgroundVitamins, Antioxidants and Endothelial Function in Coronary Artery Disease
Hornig, B. (2002) · Cardiovascular Drugs and Therapy · doi:10.1023/a:1022182201534 · Cited by 17
“It should be noted, however, that ascorbate prevents the interaction of superoxide anions and nitric oxide only at very high physiological concentrations [21] and that in all human studies only high doses of vitamin C were effective.”
BackgroundN -desethylamiodarone modulates intracellular calcium concentration in endothelial cells
Himmel, H. M., Dobrev, D., Grossmann, M., & Ravens, U. (2000) · Naunyn-Schmiedeberg s Archives of Pharmacology · doi:10.1007/s002100000301 · Cited by 16
“Since persistent micromolar Ca 2+ concentrations may be caused by reactive oxygen species or free radicals (Geeraerts et al. 1991; Volk et al. 1997; for review see Kourie 1998), we examined the effect of the antioxidant ascorbic acid (Frei et al. 1989; Jackson et al. 1998 ).”
BackgroundSerra, P. A., Esposito, G., Delogu, M. R., Migheli, R., Rocchitta, G., MIELE, E., Desole, M. S., & Miele, M. (2001) · British Journal of Pharmacology · doi:10.1038/sj.bjp.0703887 · Cited by 16
“Ascorbic acid may either generate NO from nitrite ions (NO2 7) in the extracellular space (Millar, 1995), protect NO from destruction by superoxide anions (Dudgeon et al., 1998; Jackson et al., 1998), or scavenge it (Whiteman & Halliwell, 1996).”
BackgroundSerra, P. A., Rocchitta, G., Esposito, G., Delogu, M. R., Migheli, R., MIELE, E., Desole, M. S., & Miele, M. (2001) · British Journal of Pharmacology · doi:10.1038/sj.bjp.0704232 · Cited by 16
“Ascorbic acid is a natural inhibitor of peroxynitrite generation (Jackson et al., 1998; Kirsch & de Groot, 2000), while uric acid is a natural scavenger of peroxynitrite (Hooper et al., 1998); however, in this study, SIN-1 infusion a ected neither ascorbic acid nor uric acid dialysate levels.”
BackgroundDehydroascorbate protection against dioxin-induced toxicity in the β-cell line INS-1E
Martino, L., Novelli, M., Masini, M., CHIMENTI, D., Piaggi, S., Masiello, P., & Tata, V. D. (2009) · Toxicology Letters · doi:10.1016/j.toxlet.2009.04.025 · Cited by 16
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“Normal plasma Vit C levels are around 50–70 μmol/L [44]; however, Vit C must reach a plasma concentration of 10 mmol/L to displace the reaction of the superoxide anion radical with nitric oxide, which acts at a rate 105 times greater than that of the reaction between ascorbic acid and the superoxide anion radical [45,…”
BackgroundModulation of Gap Junction-Dependent Arterial Relaxation by Ascorbic Acid
Edwards, D. H., Chaytor, A. T., Bakker, L. M., & Griffith, T. M. (2007) · Journal of Vascular Research · doi:10.1159/000104254 · Cited by 15
“It is also conceivable that AA could modulate the effects of residual NO activity, since there is evidence that the vitamin can either decrease or increase the bioavailability of NO according to experimental conditions [ , , , ,”
BackgroundHigh-Dose Intravenous Ascorbic Acid: Ready for Prime Time in Traumatic Brain Injury?
Leichtle, S. W., Sarma, A. K., Strein, M., Yajnik, V., Rivet, D. J., Sima, A. P., & Brophy, G. M. (2019) · Neurocritical Care · doi:10.1007/s12028-019-00829-x · Cited by 16
Yang, M., Bhopale, V. M., & Thom, S. R. (2015) · Journal of Applied Physiology · doi:10.1152/japplphysiol.00183.2015 · Cited by 15
“These events are inhibited by ascorbic acid, which is an efficient singlet O2 scavenger but a relatively poor scavenger for more highly reactive oxygen species, nitric oxide (NO·), and related agents such as peroxynitrite (5, 19).”
BackgroundPharmacological Approaches to the Treatment of Oxidative Stress-Induced Cardiovascular Dysfunctions
Zamora, P. L., & Villamena, F. A. (2013) · Future Medicinal Chemistry · doi:10.4155/fmc.13.15 · Cited by 14
Sicard, P., Oudot, A., Guilland, J.-C., Moreau, D., Vergely, C., & Rochette, L. (2006) · Vascular Pharmacology · doi:10.1016/j.vph.2006.04.001 · Cited by 15
“Indeed, physiologic concentrations of vitamin C are unlikely to play a role in countering the interaction of nitric oxide and superoxide anion (Jackson et al., 1998), a mechanism that has been put forward to support the oxidative hypothesis of hypertension.”
BackgroundRedox modulation of the inotropic response to dobutamine is impaired in patients with heart failure
Mak, S. S., & Newton, G. E. (2004) · American Journal of Physiology-Heart and Circulatory Physiology · doi:10.1152/ajpheart.00633.2003 · Cited by 15
“It has been demonstrated that vitamin C effectively suppresses lipid peroxidation in human plasma (10) and that vitamin C is an effective scavenger of the superoxide anion radical (20), an important ROS in aerobic organisms.”
BackgroundDiscovery of Water‐Soluble Antioxidant Flavonols without Vasorelaxant Activity
Yap, S., Loft, K. J., Woodman, O. L., & Williams, S. J. (2008) · ChemMedChem · doi:10.1002/cmdc.200800146 · Cited by 15
Divergent effects of vitamin C on relaxations of rabbit aortic rings to acetylcholine and NO‐donors
de Saram, K., McNeill, K. L., Khokher, S., Ritter, J. M., & Chowienczyk, P. (2002) · British Journal of Pharmacology · doi:10.1038/sj.bjp.0704541 · Cited by 15
“However, the rate constant for the reaction of NO with O2 7 greatly exceeds that for the reaction of ascorbate with O2 7 making this mechanism unlikely except possibly at ascorbate concentrations achieved during intra-arterial delivery of vitamin C (Jackson et al., 1998).”
BackgroundParadoxical effects of vitamin C in Chagas disease
Castanheira, J. R. P. T., Castanho, R. E. P., Rocha, H., Pagliari, C., Duarte, M. I. S., Therezo, A. L. S., Chagas, E. F. B., & Martins, L. P. A. (2018) · Parasitology International · doi:10.1016/j.parint.2018.05.002 · Cited by 13
How does ascorbate improve endothelial dysfunction? - A computational analysis
Panday, S., Kar, S., & Kavdia, M. (2021) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2021.01.031 · Cited by 14
Integrated molecular response of exposure to traffic-related pollutants in the US trucking industry
Walker, D. I., Hart, J. E., Patel, C. J., Rudel, R. A., Chu, J., Garshick, E., Pennell, K. D., Laden, F., & Jones, D. P. (2021) · Environment International · doi:10.1016/j.envint.2021.106957 · Cited by 13
“(Brown and Hu 2001; Heller et al. 2001; Jackson et al. 1998; Padayatty et al. 2003) Increased oxidation, particularly of thiol couples, and subsequent shift towards a more positive steady state redox potential has been shown to regulate early events in atherosclerosis and is associated with CVD and endothelial dysfunc…”
Mak, S. S., Overgaard, C. B., & Newton, G. E. (2005) · American Journal of Physiology-Heart and Circulatory Physiology · doi:10.1152/ajpheart.00453.2005 · Cited by 13
“Importantly, the dose of vitamin C in our studies (96 mg/min) was selected to achieve an intracoronary concentration of at least 1 mmol/l, which is sufficient in vitro to compete with O2 and prevent its participation in a reaction with, and thereby consuming, NO (9, 14, 28).”
BackgroundYap, S., Woodman, O. L., Crack, P. J., & Williams, S. J. (2011) · Bioorganic & Medicinal Chemistry Letters · doi:10.1016/j.bmcl.2011.03.040 · Cited by 13
Papel de los antioxidantes en la prevención de la enfermedad cardiovascular
Páramo, A. J., Orbe, M. A. J., & Rodríguez, J. A. (2001) · Medicina Clínica · doi:10.1016/s0025-7753(01)71926-4 · Cited by 13
Systemic and renal effect of nicotine in non-smokers
Halimi, J., & Mimran, A. (2000) · Journal of Hypertension · doi:10.1097/00004872-200018110-00019 · Cited by 13
Consumption of flavonoid-rich fruits and risk of CHD: a prospective cohort study
Yang, Y., Dong, J.-Y., Cui, R., Muraki, I., Yamagishi, K., Sawada, N., Iso, H., & Tsugane, S. (2020) · British Journal Of Nutrition · doi:10.1017/s0007114520001993 · Cited by 14
“Vitamin C can also decrease the bioavailability of superoxide, relieving superoxide-mediated nitric oxide inactivation(28).”
BackgroundRezk‐Hanna, M., Seals, D. R., Rossman, M. J., Gupta, R., Nettle, C. O., Means, A., Dobrin, D., Cheng, C., Brecht, M., Mosenifar, Z., et al. (2021) · Journal of the American Heart Association · doi:10.1161/jaha.120.019271 · Cited by 12
Vitamin C for Sepsis and Septic Shock
Ammar, M. A., Ammar, A. A., Condeni, M. S., & Bell, C. M. (2021) · American Journal of Therapeutics · doi:10.1097/mjt.0000000000001423 · Cited by 12
Fujii, N., Meade, R. D., Paull, G., McGinn, R. N., Foudil‐bey, I., Akbari, P., & Kenny, G. P. (2015) · Journal of Applied Physiology · doi:10.1152/japplphysiol.00025.2015 · Cited by 12
Oxidative Stress and Essential Hypertension
Rodrigo, R., Brito, R., & González, J. (2016) · InTech eBooks · doi:10.5772/64079 · Cited by 11
Central Infusion of L-Arginine or Superoxide Dismutase Does Not Alter Arterial Pressure in SHR.
Kagiyama, S., Tsuchihashi, T., Abe, I., Matsumura, K., & Fujishima, M. (2000) · Hypertension Research · doi:10.1291/hypres.23.339 · Cited by 12
Stone, R. M., Ainslie, P. N., Tremblay, J. C., Akins, J. D., MacLeod, D. B., Tymko, M. M., DeSouza, C. A., & Bain, A. R. (2021) · The Journal of Physiology · doi:10.1113/jp282281 · Cited by 11
Effect of ascorbic acid on forearm reactive hyperaemia in patients with hypercholesterolaemia
Protogerou, A. D., Lekakis, J. P., Kontoyanni, D. D., Stamatelopoulos, K., Tsotsoros, N. D., Papaioannou, T., Tryfonopoulos, D. J., Papamichael, C. M., & Stamatelopoulos, S. (2004) · European Journal of Cardiovascular Prevention & Rehabilitation · doi:10.1097/01.hjr.0000095049.46631.b9 · Cited by 11
Rock, C. R., Miller, S., & Allison, B. J. (2024) · Antioxidants · doi:10.3390/antiox13111400 · Cited by 10
“Jackson et al. [84] previously demonstrated that although vitamin C can scavenge superoxide at low concentrations, endothelium-mediated vasodilation only resulted from high concentrations of vitamin C.”
Rabeya, I., Meesungnoen, J., & Jay‐Gerin, J. (2025) · Antioxidants · doi:10.3390/antiox14040406 · Cited by 10
“Similarly, these radicals exhibit minimal reactivity with • NO [110].”
InfluentialKaida, S., Ohta, Y., Imai, Y., & Kawanishi, M. (2010) · Redox Report · doi:10.1179/174329210x12650506622925 · Cited by 10
Influence of physical inactivity on arterial compliance during a glucose challenge
Credeur, D. P., Reynolds, L. J., Holwerda, S. W., Vranish, J. R., Young, B. E., Wang, J., Thyfault, J. P., & Fadel, P. J. (2018) · Experimental Physiology · doi:10.1113/ep086713 · Cited by 10
Gao, L., Chong, E., Pendharkar, S. A., Phillips, A., Ke, L., Li, W., & Windsor, J. A. (2021) · Frontiers in Nutrition · doi:10.3389/fnut.2021.734558 · Cited by 10
“While direct antioxidant scavenging effects exist at markedly supraphysiologic levels (about 1–10 mmol/L, 10–100 times higher than the normal levels) (66), highlighting the use of high-dose (10–16 g/day) AA administration (65, 67, 68).”
BackgroundCoronary endothelial dysfunction is not rapidly reversible with ascorbic acid
Widlansky, M. E., Biegelsen, E. S., Hamburg, N. M., Duffy, S. J., Keaney, J. F., & Vita, J. A. (2003) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2003.10.016 · Cited by 10
Tetrahydrobiopterin in the vascular system
Walter, R. B., Schaffner, A., & Schoedon, G. (2001) · Pteridines · doi:10.1515/pteridines.2001.12.3.93 · Cited by 10
Achiri, R., Mesli, F., Benomari, F. Z., Djabou, N., Tabti, B., Muselli, A., & Dib, M. E. A. (2021) · Journal of Biomolecular Structure and Dynamics · doi:10.1080/07391102.2021.1971563 · Cited by 9
“Research by (Jackson et al., 1998) has shown that normal extracellular concentrations of ascorbic acid (30 to 150 micromoles/L) are not likely to prevent the interaction of NO (nitric oxide) with superoxide in physiological conditions.”
Background
Metadata
- Authors
- Jackson, T. S., Xu, A., Vita, J. A., & Keaney, J. F.
- Affiliations
- Boston University (US)
- Venue
- Circulation Research
- Published
- 1998-11-02
- Volume
- 83
- Issue
- 9
- Pages
- 916-922
- ISSN
- 0009-7330 (ISSN-L)
- 1524-4571
- OpenAlex
- W2155603952
- Cited by
- 400
- References
- 48
- Citations by year
- 2025: 9 · 2024: 6 · 2023: 4 · 2022: 4 · 2021: 18 · 2020: 15 · 2019: 13 · 2018: 9 · 2017: 7 · 2016: 14 · 2015: 12 · 2014: 17 · 2013: 17 · 2012: 21
- FWCI
- 11.04
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Circulation Research (publishedVersion)
- PubMed (publishedVersion)
- Funders
- American Heart Association
- National Institutes of Health (5r01hl059346-02, HL59346, HL55854, 1r01hl053398-01a1)
- National Heart, Lung, and Blood Institute (HL59346, HL 53398, P01HL055854, HL-55854)
- Topics
- Health Sciences › Medicine › Physiology (medical) › Nitric Oxide and Endothelin Effects
- Health Sciences › Medicine › Biochemistry (medical) › Antioxidant Activity and Oxidative Stress
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Keywords
- vitamin C, superoxide, nitric oxide, superoxide dismutase, peroxynitrite, nitrotyrosine, endothelium-derived nitric oxide, acetylcholine-induced relaxation, diethyldithiocarbamate
- MeSH terms
- Animals, Ascorbic Acid, Endothelium, Vascular, Female, Male, Nitric Oxide, Pyrogallol, Rabbits, Superoxides, Vasodilation
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