Abstract
Nitric oxide (NO) acts as a regulator in cell proliferation and expression of growth factors and forms peroxynitrite (ONOO−) in oxidative conditions. The aim of the study was to investigate the role of NO in cellular response to hyperbaric oxygen (HBO). NO and nitrotyrosine (NT), biochemical marker for ONOO−, cell proliferation and growth factors, were ex-vivo studied in cell cultures under HBO and normobaric (NOR) conditions. A549 (epithelial), L929 (fibroblast) and SVEC (endothelial) were exposed to 100% O2, at P = 280 kPa for t = 60 min, once daily for five sessions. Cell proliferation was determined as the incorporation of bromodeoxyuridine (BrdU) into cells and NO as nitrates/nitrites (NO3 −/ NO2 −) Gries reaction product in cell culture supernatant (CCSP). NT, vascular endothelial growth factor (VEGF) and transforming growth factor-beta 1 (TGFb1) were measured with enzyme-inked immunosorbent assay (ELISA) in CCSP. The time course of total NO was opposite to that of cell proliferation in HBO conditions, peaking after the second HBO session, while cell proliferation showed a reverse trend, minimizing at the same time, suggesting a reverse and transient anti-proliferative effect. Released growth factors were significantly increased in late HBO sessions. NT peaked after second treatment, indicating the formation of ONOO−. In control cultures (NOR), proliferation rate was downward and no significant differences were found for the other parameters. In conclusion, the data suggested a key role for NO in the beneficial HBO action, depending on its concentration, which fluctuated with the time of HBO exposure and the activation of oxidant–antioxidant (REDOX) mechanisms, regardless of cell type.
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References
Allen BW, Demchenko IT, Piantadosi CA (2009) Two faces of nitric oxide: implications for cellular mechanisms oxygen toxicity. J Appl Physiol 106:662–667
Arnal JF, Tack I, Besombes JP, Pipy B, Negre-Salvayre A (1996) Nitric oxide and superoxide anion production during endothelial cell proliferation. Am J Physiol 271:C1521–C1526
Ay H, Topal T, Ozler M, Uysal B, Korkmaz A, Oter S, Ogur R, Dundar K (2007) Persistence of hyperbaric-induced oxidative effects after exposure in rat brain cortex tissue. Life Sci 80:2025–2029
Bemedetti S, Lamorgese A, Piersantelli M, Pagliarani S, Benvenuti F, Canestrani F (2004) Oxidative stress and antioxidant status in patients undergoing prolonged exposure to hyperbaric oxygen. Clin Biochem 37:312–317
Boykin JV Jr, Baylis C (2007) Hyperbaric oxygen therapy mediates increased nitric oxide production associated with wound healing: a preliminary study. Adv Skin Wound Care 20(7):382–388
Bryan SN, Fernadez BO, Bauer SM, Garcia-Saura MF, Milsom AB, Tienush R, Maloney RE, Bharti A, Rodriguez J, Feelisch M (2005) Nitrite is a signaling molecule and regulator of gene expression in mammalian tissues. Nat Chem Biol 1(5):291–297
Buras JA, Veves A, Orlow D, Reenstra WR (2001) The effects of hyperbaric oxygen on cellular proliferation and platelet-derived growth factor receptor expression in non-insulin-dependent diabetic dermal fibroblasts. Acad Emerg Med 8(5):518–519
Camporesi EM (1996) In: Hyperbaric oxygen therapy: a committee report. Rev. Bethesda, Undersea and Hyperbaric Medical Society
Demchenko IT, Atochin DM, Boso AE, Astern J, Huang PL, Piantadosi CA (2003) Oxygen seizure latency and peroxynitrite formation in mice lacking neuronal or endothelial nitric oxide synthases. Neurosci Lett 344:53–56
Dennog C, Hartamnn A, Frey G, Speit G (1996) Detection of DNA damage after hyperbaric oxygen (HBO) therapy. Mutagenesis 11:605–609
Dulak J, Jozkowicz A (2003) Regulation of vascular endothelial growth factor synthesis by nitric oxide: facts and controversies. Antioxid Redox Signal 5(1):123–132
Gendimenico GJ, Schlesinge HR, Ritter MA, Haugaard N (1984) Inhibition of growth and decreased survival of B104 rat neuroblastoma cells after exposure to hyperbaric oxygen. In Vitro 20(5):385–390
Gilchrist M, Shore AC, Benjamin N (2011) Inorganic nitrate and nitrite and control of blood pressure. Cardiovasc Res 89:492–498
Gill AL, Bell CAN (2004) Hyperbaric oxygen: its uses, mechanisms of action and outcomes. Q J Med 97:385–395
Granowitz EV, Tonomura N, Benson RM, Katz DM, Band V, Makari-Judson GP, Osborne BA (2005) Hyperbaric oxygen inhibits benign and malignant human mammary epithelial cell proliferation. Anticancer Res 25:3833
Groeger M, Speit G, Radermacher P, Muth CM (2005) Interaction of hyperbaric oxygen, nitric oxide, and heme oxygenase on DNA strand breaks in vivo. Mutat Res 572:167–172
Groeger M, Oter S, Simkova V, Bolten M, Koch A, Warninghoff V, Georgief M, Muth CM, Speit G, Radermacher P (2009) DNA damage after long-term repetitive hyperbaric exposure. J Appl Physiol 106(1):311–315
Havesa O, Ramosb B, Rodriguez LL, Aguilara A, Badiac T, Castro FO (2005) Cell confluency is as efficient as serum starvation for inducing arrest in the G0/G1 phase of the cell cycle in granulosa and fibroblast cells of cattle. Anim Reprod Sci 87(3):181–192
Hink J, Jansen E (2001) Are superoxide and/or hydrogen peroxide responsible for some of the beneficial effects of hyperbaric oxygen therapy? Med Hypotheses 57(6):764–769
Hink J, Thom SR, Simonsen U, Rubin I, Jansen E (2006) Vascular reactivity and endothelial NOS activity in rat thoracic aorta during and after hyperbaric oxygen exposure. Am J Physiol Heart Circ Physiol 291:1988–1998
Hoeben A, Landuyt B, Highley MS, Wildiers H, Van Oosterom AT, De Bruijn EA (2004) Vascular endothelial growth factor and angiogenesis. Pharm Rev 56(4):549–580
Huang KL, Wu GN, Lin HC, Mao SP, Kang BH, Wan FJ (2000) Prolonged exposure to hyperbaric oxygen neuronal damage in primary rat cortical cultures. Neurosci Lett 293:159–162
Huang KT, Han TH, Hyduke DR, Vaughn MW, Van Herle H, Hein TW, Zhang C, Kuo L, Liao JC (2001) Modulation of nitric oxide bioavailability by erythrocytes. Proc Natl Acad Sci USA 98 (20):11771–11776
Inoue M, Sato EF, Park AM, Nishikawa M, Kasahara E, Miyoshi M, Ochi A, Utsumi K (2000) Cross-talk between NO and oxyradicals, a supersystem that regulates energy metabolism and survival of animals. Free Radic Res 33(6):757–770
Kang TS, Gorti GK, Quan SY, Ho M, Koch RJ (2004) Effect of hyperbaric oxygen on the growth factor profile of fibroblasts. Arch Facial Plast Surg 6:31–35
Kimura H, Esumi H (2003) Reciprocal regulation between nitric oxide and vascular growth factor in angiogenesis. Acta Biochim Pol 50(1):49–59
Messier AA, Fisher HW (1990) Sensitivity of cultured mammalian cells to oxidative stress: adaptation to repeated exposures of hyperbaric oxygen. Undersea Biomed Res 17:305–322
Mizuguchi T, Oshima H, Imaizumi H, Kohara H, Kawamoto M, Nobuoka T, Kawasaki H, Harada K, Masuda Y, Kikkawa Y, Mitaka T, Asai Y, Hirata K (2005) Hyperbaric oxygen stimulates cell proliferation and normalizes multidrug resistance protein-2 protein localization in primary rat hepatocytes. Wound Rep Regen 13:551–557
Okragly A, Balwit JM, Haak-Frendscho M (1994) Transforming growth factor beta-1 (TGF-beta1): a biological paradox. Promega Notes Magazine 47:10–17
Pacher P, Beckman JS, Liaudet L (2007) Nitric oxide and peroxynitrite in health and disease. Physiol Rev 87:315–424
Papapetropoulos A, Garcia-Cardena G, Madri J, Sessa W (1997) Nitric oxide production contributes to the angiogenetic properties of vascular endothelial growth factor in human endothelial cells. J Clin Invest 100:3131–3139
Patel V, Chivukuta IV, Roy S, Khanna S, He G, Ojha N, Mehotra A, Dias LM, Hunt TK, Sen CK (2005) Oxygen: from the benefits of inducing VEGF expression to managing the risk of hyperbaric stress. Antioxid Redox Signal 7(9–10):1377–1387
Russmann E, Sieber K, Doppler C, Hinzpeter M (1993) Cell proliferation ELISA, BrdU (chemiluminescence). In: Coligan JE et al (eds) Current protocols in immunology, vol 1. Wiley, New York. Colloquium Roche Molecular Biochemicals 4:1–4
Shackelford RE, Kaufmann WK, Paulis RS (2000) Oxidative stress and cell cycle check point function. Free Rad Biol Med 28(9):1387–1404
Speit G, Bonzheim I (2003) Genotoxic and protective effects of hyperbaric oxygen in A549 lung cells. Mutagenesis 18(6):545–548
Strober W (1997) Trypan blue exclusion test of cell viability. Current protocols in immunology. A.3B.1–A.3B.2 Copyright © 1997, Wiley
Tompach PC, Lew D, Stoll JL (1997) Cell response to hyperbaric treatment. Int J Oral Maxillofac Surg 26:82–86
Tsikas D (2005) Methods of quantitative analysis of the nitric oxide metabolites nitrite and nitrate in human biological fluids. Free Radic Res 39:797–815
Vega CJ, Peterson DA (2005) Stem cell proliferative history in tissue revealed by temporal halogenated thymidine analog discrimination. Nat Methods 2:167–169
Villalobo A (2006) Nitric oxide and cell proliferation. FEBS J 273:2329–2344
Xu X, Yi H, Kato M, Suzuki H, Kobayasho S, Takahashi H, Nakashima I (1997) Differential sensitivities to hyperbaric oxygen of lymphocyte subpopulations of normal and autoimmune mice. Immunol Lett 59(2):79–84
Acknowledgments
We thank Professor J. I. Sznajder (Northwestern University, Chicago USA), Professor Sadis Matalon (University of Alabama at Birmingham, USA), Professor C. Sekeris (Medical School, University of Athens, Greece) and Professor A. Papapetropoulos (Nursing School University of Athens, Greece) for the kindly offered cell lines. The project was provided a partial grant from the Athens University and supported by the KAT Hospital.
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Communicated by Susan A. Ward.
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Venetsanou, K., Fildissis, G., Tokta, R. et al. The role of nitric oxide in cellular response to hyperbaric conditions. Eur J Appl Physiol 112, 677–687 (2012). https://doi.org/10.1007/s00421-011-2027-8
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DOI: https://doi.org/10.1007/s00421-011-2027-8