Abstract
Reactive oxygen species (ROS) play a pivotal role in UVA-induced cell damage. As expression of the inducible nitric oxide synthase (iNOS) is a normal response of human skin to UV radiation we examined the role of nitric oxide (NO) as a protective agent during or even after UVA1- or ROS-exposure against apoptosis or necrosis of rat endothelial cells. When added during or up to 2 h subsequent to UVA1 or ROS exposure the NO-donor S-nitroso-cysteine (SNOC) at concentrations from 100–1000 μM significantly protects from both apoptosis as well as necrosis. The NO-mediated protection strongly correlates with complete inhibition of lipid peroxidation (sixfold increase of malonedialdehyde formation in untreated versus 1.2-fold with 1 mM SNOC). NO-mediated protection of membrane function was also shown by the inhibition of cytochrome c leakage in UVA1 treated cells, a process not accompanied by alterations in Bax and Bcl-2 protein levels. Thus, the experiments presented demonstrate that NO exposure during or even after a ROS-mediated toxic insult fully protects from apoptosis or necrosis by maintaining membrane integrity and function.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- BHT:
-
butylated hydroxytoluene
- EC:
-
endothelial cells
- iNOS:
-
inducible nitric oxide synthase
- NO:
-
nitric oxide
- RB/hν:
-
Rose Bengal plus visible light
- ROS:
-
reactive oxygen species
- SNOC:
-
S-nitrosocysteine
- SOD:
-
superoxide dismutase
- UVA1:
-
ultraviolet radiation A1
References
Passchier WF, Bosnjakovic BFM . 1987 Human Exposure to Ultraviolet Radiation. Amsterdam
Webb RB . 1977 Lethal and mutagenic effects of near-ultraviolet radiation. In Photochemistry and Photobiology Reviews. Smith KC, ed New York: Plenum pp. 169–262
Tyrrell RM, Keyse SM . 1990 The interaction of UVA radiation with cultured cells J. Photochem. Photobiol. 4B: 349–361
Black HS . 1987 Potential involvement of free radical reactions in ultraviolet light-mediated cutaneous damage Photochem. Photobiol. 46: 213–221
Adams JM, Cory S . 1998 The Bcl-2 protein family: arbiters of cell survival Science 281: 1322–1326
Kim Y-M, Bombeck CA, Billiar TR . 1999 Nitric oxide as a bifunctional regulator of apoptosis Circ. Res. 84: 253–256
Förstermann U, Closs EI, Pollock JS, Nakane M, Schwarz P, Gath I, Kleinert H . 1994 Nitric oxide synthase isoenzymes. Characterisation, purification, molecular cloning, and functions Hypertension 23: 1121–1131
Kröncke K-D, Fehsel K, Kolb-Bachofen V . 1995 Inducible nitric oxide synthase and its product nitric oxide, a small molecule with complex biological activities Biol. Chem. 376: 327–343
Suschek C, Rothe H, Fehsel K, Enczmann J, Kolb-Bachofen V . 1993 Induction of a macrophage-like nitric oxide synthase in cultured rat aortic endothelial cells J. Immunol. 151: 3283–3291
Suschek CV, Krischel V, Bruch-Gerharz D, Berendji D, Krutmann J, Kröncke K-D, Kolb-Bachofen V . 1999 Nitric oxide fully protects against UVA-induced apoptosis in tight correlation with Bcl-2 up-regulation J. Biol. Chem. 274: 6130–6137
Klotz L-O, Pellieux C, Briviba K, Pierlot C, Aubry J-M, Sies H . 1999 Mitogen-activated protein kinase (p38, JNK-, ERK-) activation pattern induced by extracellular and intracellular singlet oxygen and UVA Eur. J. Biochem. 260: 917–922
Briviba K, Klotz L-O, Sies H . 1997 Toxic and signaling effects of photochemically or chemically generated singlet oxygen in biological systems Biol. Chem. 378: 1259–1265
Tyrrell RM, Pidoux M . 1989 Singlet oxygen involvement in the inactivation of cultured human fibroblasts by UVA (334 nm, 365 nm) and near-visible (405 nm) radiation Photochem. Photobiol. 49: 407–412
Crow JP . 1997 Dichlorodihydrofluorescein and dihydrorhodamine 123 are sensitive indicators of peroxynitrite in vitro: implications for intracellular measurement of reactive nitrogen and oxygen species Nitric Oxide 1: 145–157
Danpure HJ, Tyrrell RM . 1976 Oxygen dependence of near-UV (365 nm) lethality and the interaction of near-UV and X-rays in two mammalian cell lines Photochem. Photobiol. 23: 171–177
McCormick JP, Fischer JR, Pachlatko JP . 1976 Characterization of a cell-lethal product from the photooxidation of tryptophan: hydrogen peroxide Science 191: 468–469
Czochralska B, Kamczynski W, Bartosz G, Shugar D . 1984 Oxidation of excited-state NADH and NAD dimer in aqueous medium involvement of O2− as a mediator in the presence of oxygen Biochim. Biophys. Acta. 801: 403–409
Peak JG, Peak MJ, Foote CS . 1982 Effects of glycerol upon the biological action of near-ultraviolet light: spectra and concentration dependence for transforming DNA and for Escherichia coli B/r Photochem. Photobiol. 3: 413–416
Kanofsky JR . 1989 Singlet oxygen production by biological systems Chem-Biol. Interact. 70: 1–28
Janssen YMW, Van Houten B, Borm PJA, Mossman BT . 1993 Biology of Disease: cell and tissue responses to oxidative damage Lab. Invest. 69: 261–274
Sies H . 1986 Biochemistry of oxidative stress Angew. Chem. Int. Ed. 25: 1058–1071
Ramakrishnan N, McClain DE, Catravas GN . 1993 Membranes as sensitive targets in thymocyte apoptosis Int. J. Radiat. Biol. 63: 693–701
Forrest VJ, Kang Y-H, McClain DE . 1994 Oxidative stress-induced apoptosis prevented by Trolox Free Radical Biol. Med. 16: 675–684
Kuhn A, Fehsel K, Lehmann P, Krutmann J, Ruzicka T, Kolb-Bachofen V . 1998 Aberrant timing in epidermal expression of inducible nitric oxide synthase after UV-irradiation in Lupus patients J. Invest. Dermatol. 111: 149–153
Leist M, Single B, Castoldi AF, Kühnle P, Nicotera P . 1997 Intracellular adenosine triphosphate (ATP) concentrations: A switch in the decision between apoptosis and necrosis J. Exp. Med. 185: 1481–1486
Kröncke K-D, Fehsel K, Kolb-Bachofen V . 1997 Nitric oxide: cytotoxicity versus cytoprotection: How, why, when, and where? Nitric Oxide Biol. Chem. 1: 107–120
Ryter SW, Tyrrell RM . 1998 Singlet molecular oxygen ((1)O2): a possible effector of eukaryotic gene expression Free Radic Biol. Med. 24: 1520–1534
McCaughan Jr JS . 1999 Photodynamic therapy: a review Drugs Aging 15: 49–68
Fukuto JM, Cho JY, Switzer CH . 2000 The chemical properties of nitric oxide and related nitrogen oxides. In: Nitric Oxide: Biology and Pathobiology. Ignarro LJ, ed San Diego: Academic Press pp. 23–40
Darley-Usmar VM, Patel RP, O'Donnell VB, Freeman BA . 2000 Antioxidant actions of nitric oxide. In: Nitric Oxide: Biology and Pathobiology. Ignarro LJ, ed San Diego: Academic Press pp. 265–276
Miranda KM, Espey MG, Jourd'heuil D, Grisham MB, Fukuto J, Feelisch M, Wink DA . 2000 The chemical biology of nitric oxide. In: Nitric Oxide: Biology and Pathobiology. Ignarro LJ, ed San Diego: Academic Press pp. 41–56
Di Mascio P, Sies H . 1989 Quantification of singlet oxygen generated by thermolysis of 3,3′-(1,4-naphthylidene)dipropionate. Monomol and dimol photoemission and the effects of 1,4-diazabicyclo [2.2.2] octane J. Am. Chem. Soc. 111: 2909–2914
Kröncke K-D, Kolb-Bachofen V . 1996 Detection of nitric oxide interaction with zinc finger proteins Meth. Enzymol. 269: 279–284
Hrabie JA, Klose JR, Wink DA, Keefer LK . 1993 New nitric oxide-releasing zwitterions derived from polyamines J. Org. Chem. 58: 1472–1476
Suschek C, Fehsel K, Kröncke K-D, Sommer A, Kolb-Bachofen V . 1994 Primary cultures of rat capillary endothelial cells: constitutive and cytokine-inducible macrophage-like nitric oxide synthases are expressed and activities regulated by glucose concentration Am. J. Pathol. 145: 485–695
Klotz L-O, Briviba K, Sies H . 1997 Singlet oxygen mediates the activation of JNK by UVA radiation in human skin fibroblasts FEBS Lett. 408: 289–291
Fehsel K, Kolb-Bachofen V, Kolb H . 1991 Analysis of TNF-alpha-induced DNA strand breaks at the single cell level Am. J. Path. 139: 251–157
Kaiser S, DiMascio P, Murphy ME, Sies H . 1990 Physical and chemical scavenging of singlet molecular oxygen by tocopherols Arch. Biochem. Biophys. 277: 101–108
Egorov SY, Kurella EG, Boldyrev AA, Krasnovsky AAJ . 1997 Quenching of singlet molecular oxygen by carnosine and related antioxidants. Monitoring 1270-nm phosphorescence in aqueous media Biochem. Mol. Biol. Int. 41: 687–694
Michaeli A, Feitelson J . 1994 Reactivity of singlet oxygen toward amino acids and peptides Photochem. Photobiol. 59: 284–289
Faulkner KM, Stevens RD, Fridovich I . 1994 Characterization of Mn(III) complexes of linear and cyclic desferrioxamines as mimics of superoxide dismutase activity Arch. Biochem. Biophys. 310: 341–346
Faulkner KM, Liochev SI, Fridovich I . 1994 Stable Mn(III) porphyrins mimic superoxide dismutase in vitro and substitute for it in vivo J. Biol. Chem. 269: 23471–23476
Heikkila RE, Cabbat FS, Cohen G . 1976 In vivo inhibition of superoxide dismutase in mice by diethyldithiocarbamate J. Biol. Chem. 251: 2182–2185
Williams RN, Delamere NA, Paterson CA . 1985 Inactivation of catalase with 3-amino-1,2,4-triazole: an indirect irreversible mechanism Biochem. Pharmacol. 34: 3386–3389
Bose B, Agarwal S, Chatterjee SN . 1989 UV-A induced lipid peroxidation in liposomal membrane Radiat. Environ. Biophys. 28: 59–65
Babbs CF, Griffin DW . 1989 Scatchard analysis of methane sulfinic acid production from dimethyl sulfoxide: a method to quantify hydroxyl radical formation in physiologic systems Free Radic. Biol. Med. 6: 493–503
Chaturvedi V, Wong B, Newman SL . 1996 Oxidative killing of Cryptococcus neoformans by human neutrophils. Evidence that fungal mannitol protects by scavenging reactive oxygen intermediates J. Immunol. 156: 3836–3840
Royall JA, Ischiropoulos H . 1993 Evaluation of 2′,7′-dihydrofluorescin and dihydrorhodamine 123 as fluorescent probes for intracellular H2O2 in cultured endothelial cells Arch. Biochem. Biophys. 302: 348–355
Knight JA, Pieper RK, McClellan L . 1988 Specificity of the thiobarbituric acid reaction: its use in studies of lipid peroxidation Clin. Chem. 34: 2433–2438
Scheschonka A, Murphy ME, Sies H . 1990 Temporal relationships between the loss of vitamin E, protein sulfhydryls and lipid peroxidation in microsomes challenged with different prooxidants Chem-Biol Interact. 74: 233–252
Acknowledgements
We thank Christa-Maria Wilkens-Roth, Annette Reimann, Marija Lenzen, and Ulla Lammersen for technical assistance, and Martha Turken for photographic assistance. This study was supported by grants from the Deutsche Forschungsgemeinschaft (SFB 503, A3 and B1) and by NFCR.
Author information
Authors and Affiliations
Corresponding author
Additional information
Edited by A Finazzi-Agro
Rights and permissions
About this article
Cite this article
Suschek, C., Briviba, K., Bruch-Gerharz, D. et al. Even after UVA-exposure will nitric oxide protect cells from reactive oxygen intermediate-mediated apoptosis and necrosis. Cell Death Differ 8, 515–527 (2001). https://doi.org/10.1038/sj.cdd.4400839
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/sj.cdd.4400839
Keywords
This article is cited by
-
Corneal endothelial loss after crosslinking with riboflavin and ultraviolet-A
Graefe's Archive for Clinical and Experimental Ophthalmology (2012)
-
Photolytically generated nitric oxide inhibits caspase activity and results in AIF-mediated cell death
Journal of Molecular Medicine (2010)
-
Topically Applied Nitric Oxide Induces T-Lymphocyte Infiltration in Human Skin, but Minimal Inflammation
Journal of Investigative Dermatology (2008)
-
Evidence for Hydroxyl Radical Scavenging Action of Nitric Oxide Donors in the Protection Against 1-Methyl-4-phenylpyridinium-induced Neurotoxicity in Rats
Neurochemical Research (2008)
-
Evaluation of a synthetic CArG promoter for nitric oxide synthase gene therapy of cancer
Gene Therapy (2005)
