Near infrared light protects cardiomyocytes from hypoxia and reoxygenation injury by a nitric oxide dependent mechanism.

Photobiomodulation with near infrared light (NIR) provides cellular protection in various disease models. Previously, infrared light emitted by a low-energy laser has been shown to significantly improve recovery from ischemic injury of the canine heart. The goal of this investigation was to test the hypothesis that NIR (670 nm) from light emitting diodes produces cellular protection against hypoxia and reoxygenation-induced cardiomyocyte injury. Additionally, nitric oxide (NO) was investigated as a potential cellular mediator of NIR. Our results demonstrate that exposure to NIR at the time of reoxygenation protects neonatal rat cardiomyocytes and HL-1 cells from injury, as assessed by lactate dehydrogenase release and MTT assay. Similarly, indices of apoptosis, including caspase 3 activity, annexin binding and the release of cytochrome c from mitochondria into the cytosol, were decreased after NIR treatment. NIR increased NO in cardiomyocytes, and the protective effect of NIR was completely reversed by the NO scavengers carboxy-PTIO and oxyhemoglobin, but only partially blocked by the NO synthase (NOS) inhibitor L-NMMA. Mitochondrial metabolism, measured by ATP synthase activity, was increased by NIR, and NO-induced inhibition of oxygen consumption with substrates for complex I or complex IV was reversed by exposure to NIR. Taken together these data provide evidence for protection against hypoxia and reoxygenation injury in cardiomyocytes by NIR in a manner that is dependent upon NO derived from NOS and non-NOS sources.

[1]  C. Richter,et al.  Nitric oxide synthase activity in mitochondria , 1997, FEBS letters.

[2]  Steven P Jones,et al.  The ubiquitous role of nitric oxide in cardioprotection. , 2006, Journal of molecular and cellular cardiology.

[3]  Dinh Tuan Vo Biomedical photonics handbook , 2003 .

[4]  C. Giulivi,et al.  Nitric oxide regulation of mitochondrial oxygen consumption I: cellular physiology. , 2006, American journal of physiology. Cell physiology.

[5]  S Nioka,et al.  Comparison of time-resolved and -unresolved measurements of deoxyhemoglobin in brain. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[6]  L. Szweda,et al.  Cardiac ischemia/reperfusion, aging, and redox-dependent alterations in mitochondrial function. , 2003, Archives of biochemistry and biophysics.

[7]  C. Cooper,et al.  Nitric oxide inhibition of respiration involves both competitive (heme) and noncompetitive (copper) binding to cytochrome c oxidase. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[8]  P. Brookes,et al.  Mitochondria as a target for the cardioprotective effects of nitric oxide in ischemia-reperfusion injury. , 2008, Antioxidants & redox signaling.

[9]  I. M. Catalano,et al.  Increase of proton electrochemical potential and ATP synthesis in rat liver mitochondria irradiated in vitro by helium‐neon laser , 1984, FEBS letters.

[10]  Nick Lane,et al.  Power games , 2006 .

[11]  U. Oron,et al.  Promotion of Bone Repair in the Cortical Bone of the Tibia in Rats by Low Energy Laser (He-Ne) Irradiation , 1996, Calcified Tissue International.

[12]  E. Jacobs,et al.  Nitric oxide contributes to 20-HETE-induced relaxation of pulmonary arteries. , 2002, Journal of applied physiology.

[13]  Noel T. Whelan,et al.  Therapeutic photobiomodulation for methanol-induced retinal toxicity , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[14]  M. Gladwin,et al.  Deoxymyoglobin Is a Nitrite Reductase That Generates Nitric Oxide and Regulates Mitochondrial Respiration , 2007, Circulation research.

[15]  C. Szabó,et al.  Mitochondrial nitric oxide synthase is not eNOS, nNOS or iNOS. , 2003, Free radical biology & medicine.

[16]  Guiding principles for research involving animals and human beings. , 2001, American journal of physiology. Regulatory, integrative and comparative physiology.

[17]  T. Karu,et al.  Irradiation with He-Ne laser increases ATP level in cells cultivated in vitro. , 1995, Journal of photochemistry and photobiology. B, Biology.

[18]  Noel T. Whelan,et al.  Effect of NASA light-emitting diode irradiation on wound healing. , 2001, Journal of clinical laser medicine & surgery.

[19]  Tuan Vo-Dinh,et al.  Biomedical Photonics Handbook , 2003 .

[20]  C. Cooper,et al.  Nitric oxide and iron proteins. , 1999, Biochimica et biophysica acta.

[21]  P. Pagel,et al.  Isoflurane Protects against Myocardial Infarction during Early Reperfusion by Activation of Phosphatidylinositol-3-Kinase Signal Transduction: Evidence for Anesthetic-induced Postconditioning in Rabbits , 2005, Anesthesiology.

[22]  G. Burnstock,et al.  Immunocytochemical evidence for a mitochondrially located nitric oxide synthase in brain and liver. , 1995, Biochemical and biophysical research communications.

[23]  K. Kawahara,et al.  Ischemia/reperfusion-induced death of cardiac myocytes: possible involvement of nitric oxide in the coordination of ATP supply and demand during ischemia. , 2006, Journal of molecular and cellular cardiology.

[24]  L. Argaud,et al.  Postconditioning Inhibits Mitochondrial Permeability Transition , 2005, Circulation.

[25]  B. Brüne The intimate relation between nitric oxide and superoxide in apoptosis and cell survival. , 2005, Antioxidants & redox signaling.

[26]  M. Gladwin,et al.  Nitrite reductase activity of myoglobin regulates respiration and cellular viability in myocardial ischemia-reperfusion injury , 2008, Proceedings of the National Academy of Sciences.

[27]  Raymond J. Lanzafame,et al.  Photomodulation of Oxidative Metabolism and Electron Chain Enzymes in Rat Liver Mitochondria , 1997, Photochemistry and photobiology.

[28]  P. Pratt,et al.  Bioassay of an Endothelium-Derived Hyperpolarizing Factor from Bovine Coronary Arteries: Role of a Cytochrome P450 Metabolite , 1998, Journal of Vascular Research.

[29]  R. Shofti,et al.  Low-Energy Laser Irradiation Reduces Formation of Scar Tissue After Myocardial Infarction in Rats and Dogs , 2001, Circulation.

[30]  Z. Bosnjak,et al.  Isoflurane preconditioning uncouples mitochondria and protects against hypoxia-reoxygenation. , 2007, American journal of physiology. Cell physiology.

[31]  T. Karu,et al.  Primary and secondary mechanisms of action of visible to near-IR radiation on cells. , 1999, Journal of photochemistry and photobiology. B, Biology.

[32]  P. Nicotera,et al.  Inhibition of mitochondrial ATP generation by nitric oxide switches apoptosis to necrosis. , 1999, Experimental cell research.

[33]  Michael Belkin,et al.  Temporal parameters of low energy laser irradiation for optimal delay of post-traumatic degeneration of rat optic nerve , 1989, Brain Research.

[34]  A. Boveris,et al.  Heart mitochondrial nitric oxide synthase. Effects of hypoxia and aging. , 2004, Molecular aspects of medicine.

[35]  P. Pagel,et al.  Cardioprotection by volatile anesthetics. , 2005, Vascular pharmacology.

[36]  E. Murphy,et al.  Preconditioning: the mitochondrial connection. , 2007, Annual review of physiology.

[37]  C M Cobb,et al.  Biostimulation of wound healing by low-energy laser irradiation. A review. , 1996, Journal of clinical periodontology.

[38]  A. Rowan Guide for the Care and Use of Laboratory Animals , 1979 .

[39]  Uri Oron,et al.  Modulations of VEGF and iNOS in the rat heart by low level laser therapy are associated with cardioprotection and enhanced angiogenesis , 2006, Lasers in surgery and medicine.

[40]  E. Jacobs,et al.  Multiple antiapoptotic targets of the PI3K/Akt survival pathway are activated by epoxyeicosatrienoic acids to protect cardiomyocytes from hypoxia/anoxia. , 2008, American journal of physiology. Heart and circulatory physiology.

[41]  N J Izzo,et al.  HL-1 cells: a cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult cardiomyocyte. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[42]  Uri Oron,et al.  Photoengineering of tissue repair in skeletal and cardiac muscles. , 2006, Photomedicine and laser surgery.

[43]  U. Oron,et al.  Enhancement of muscle regeneration in the rat gastrocnemius muscle by low energy laser irradiation , 1992, Anatomy and Embryology.

[44]  M. Wong-Riley,et al.  Light-emitting diode treatment reverses the effect of TTX on cytochrome oxidase in neurons , 2001, Neuroreport.

[45]  Britton Chance,et al.  Photobiomodulation Directly Benefits Primary Neurons Functionally Inactivated by Toxins , 2005, Journal of Biological Chemistry.

[46]  K. Kawahara,et al.  Increased Resistance to Nitric Oxide Cytotoxicity Associated with Differentiation of Neuroblastoma-Glioma Hybrid (NG108-15) Cells , 2002, Free radical research.

[47]  S. Srinivasula,et al.  Cytochrome c and dATP-Dependent Formation of Apaf-1/Caspase-9 Complex Initiates an Apoptotic Protease Cascade , 1997, Cell.

[48]  E. Giordano,et al.  Nitric oxide synthase activity in rat cardiac mitochondria , 2004, Basic Research in Cardiology.

[49]  R. Balaban,et al.  Nitric oxide synthase in porcine heart mitochondria: evidence for low physiological activity. , 2001, American journal of physiology. Heart and circulatory physiology.

[50]  A. Landar,et al.  Nitroxia: the pathological consequence of dysfunction in the nitric oxide-cytochrome c oxidase signaling pathway. , 2005, Free radical biology & medicine.

[51]  J. Vinten-johansen,et al.  Postconditioning: reduction of reperfusion-induced injury. , 2006, Cardiovascular research.

[52]  L. Hagenfeldt,et al.  Measurement of ATP production and respiratory chain enzyme activities in mitochondria isolated from small muscle biopsy samples. , 2002, Analytical biochemistry.

[53]  J. Klein,et al.  Metallothionein inhibits doxorubicin-induced mitochondrial cytochrome c release and caspase-3 activation in cardiomyocytes. , 2001, The Journal of pharmacology and experimental therapeutics.

[54]  H. Fliss,et al.  Apoptosis in ischemic and reperfused rat myocardium. , 1996, Circulation research.

[55]  P. Anversa,et al.  Acute myocardial infarction in humans is associated with activation of programmed myocyte cell death in the surviving portion of the heart. , 1996, Journal of molecular and cellular cardiology.

[56]  E. Antonini,et al.  Studies on the relations between molecular and functional properties of hemoglobin. I. The effect of salts on the molecular weight of human hemoglobin. , 1961, The Journal of biological chemistry.

[57]  E. Solary,et al.  Imaging of nitric oxide in a living vertebrate using a diamino-fluorescein probe. , 2007, Free radical biology & medicine.

[58]  M. Duchen,et al.  Effects of NO on mitochondrial function in cardiomyocytes: Pathophysiological relevance. , 2006, Cardiovascular research.

[59]  U. Oron,et al.  Attenuation of the process of muscle regeneration in the toad gastrocnemius muscle by low energy laser irradiation , 1994, Lasers in surgery and medicine.

[60]  Yasutaka Nakamura,et al.  Low Power Laser Therapy , 1986 .

[61]  Tiina I. Karu,et al.  Irradiation with HeNe laser increases ATP level in cells cultivated in vitro , 1995 .

[62]  R. Kitsis,et al.  A mechanistic role for cardiac myocyte apoptosis in heart failure. , 2003, The Journal of clinical investigation.

[63]  J. Weiss,et al.  Mitochondria and Ischemia/Reperfusion Injury , 2005, Annals of the New York Academy of Sciences.

[64]  U. Oron,et al.  Promotion of muscle regeneration in the toad (Bufo viridis) gastrocnemius muscle by low‐energy laser irradiation , 1993, The Anatomical record.

[65]  Bernhard Brüne,et al.  Nitric oxide: NO apoptosis or turning it ON? , 2003, Cell Death and Differentiation.

[66]  C A Beltrami,et al.  Apoptosis in the failing human heart. , 1997, The New England journal of medicine.

[67]  J. Balligand Regulation of cardiac beta-adrenergic response by nitric oxide. , 1999, Cardiovascular research.

[68]  H. Toyokawa,et al.  Promotive Effects of Far-Infrared Ray on Full-Thickness Skin Wound Healing in Rats , 2003, Experimental biology and medicine.

[69]  R. Kloner,et al.  Reperfusion injury induces apoptosis in rabbit cardiomyocytes. , 1994, The Journal of clinical investigation.