ENERGETIC AND ANTIOXIDANT STATUS OF RAT LIVER MITOCHONDRIA DURING HYPOXIA−REOXYGENATION OF DIFFERENT DURATION
O.A. Gonchar1, V.I Nosar1, L.V. Bratus1, I.N. Туmchenko2, N.N. Steshenko1, I.N. Mankovska1
- O.O.Bogomoletz Institute of Physiology NAS of Ukraine, Kyiv;
- Kyiv Medical University of UAFM
Dynamics of changes in activity and protein expression of
antiradical (MnSOD), glutathione-dependent (glutathione
peroxidase, glutathione reductase) and NADP+-generated
(isocitrate dehydrogenase) enzymes as well as in the energy
metabolism indeces in rat liver mitochondria under hypoxia−
reoxygenation of different duration (1, 3, 7 14 days) were
studied. Prolonged hypoxia− reoxygenation was characterized
by phase changes of the corticosterone concentration in
rat blood, which corresponded to the changes in energy
metabolism as well as in pro− and antioxidant balance in rat
liver mitochondria. It has been shown that short−term (1day)
hypoxia− reoxygenation (5% O2 in the gas mixture) led to
an increase in the blood corticosterone concentration and
a significant activation of oxidative processes and energy
metabolism in rat liver mitochondria, the intensity of which
was reduced to 3rd day. Long− term hypoxia − reoxygenation
(7−14 th days) led to the gradual depletion of the organism
adaptive capabilities, as evidenced by a significant decline
in the blood corticosterone concentration, an increase in
the content of secondary products of lipid peroxidation, an
imbalance in pro− and antioxidant reactions and reduction
of energy capacity in liver cells mitochondria. It has been
shown that the glutathione peroxidase protein expression
and enzymatic activity increased constantly during the whole
experimental period and correlated positively with the level of
H2O2. The amount of Mn-SOD protein as well as it’s enzymatic
activity was lower in the first seven days of experiment, and it
was increased in consequent days up to the control level on 14th
day. Increased activity of glutathione peroxidase, glutathione
reductase and NADP+-dependent isocitrate dehydrogenase
during prolonged hypoxia − reoxygenation indicates that
glutathione- and NADPH-generating enzymes, were actively
involved in the antioxidant protect.
hypoxia−reoxygenation; mitochondria; antioxidative enzymes; protein expression.
- Li C, Jackson RM. Reactive species mechanisms of cellular hypoxia-reoxygenation injury. Am J Physiol. 2002 ;282 (Cell Physiol.): C227-C241.
- Baraboy VA., Reznikov OG. Physiology, biochemistry and psychology of stress. Kyiv: Interservis; 2013. [Ukrainian].
- Meerson FZ, Pshennikova MG. Adaptation to the stress situations and to physical loads. Moscow: Medicina;1988. [Russian].
- Jezek P, Hlavata L. Mitochondria in homeostasis of reactive oxygen species in cell, tissues, and organism. Int J Biochem & Cell Biol. 2005; 37: 2478-2503.
- Sazontova TG, Anchidhkina NA, Zhukova AG, Bedareva IV, Pylaeva EA, et al. Active oxygen forms and their redox-signaling role in adaptation to oxygen contents changing. Fisiol J. 2008; 2: 18-32. [Russian].
- Menshikova EV, Zenkov NK. Antioxidants and Inhibitors of radical oxidative processes. Usp Sovr Biol. 1993;113(4): 442-53. [Russian].
- Limon-Pacheco J, Gonsebatt M. The role of antioxidants and antioxidant-related enzymes in protective responses to environmentally induced oxidative stress. Mutat Res. 2009; 674:137-47.
- Hayes J, McLellan L. Glutathione and glutathione-dependent enzymes represent a coordinately regulated defense against oxidative stress. Free Rad Res. 1999; 31: 273-300.
- Jo S, Son M, Koh H, Lee S, Song I, et al. Control of mitochondrial redox balance and cellular defense against oxidative damage by mitochondrial NADP+-dependent isocitrate dehydrogenase. J Biol Chem. 2001;276: 16168-176.
- Clanton TL, Klawitter PF. Adaptive responses of skeletal muscle to intermittent hypoxia: the known and the unknown. J Appl Physiol. 2007; 90: 2476-87.
- Gonchar O, Mankovskaya I. Effect of moderate hypoxia/ reoxygenetion on mitochondrial adaptation to acute severe hypoxia. Acta Biol Hung. 2009; 60: 185-94.
- Lukyanova LD. Novel approach to the understanding of molecular mechanisms of adaptation to hypoxia. In: Adaptation Biology and Medicine, eds Hargens A, Takeda N, Singal PCurrent Concepts, New Delhi: Narosa 2005; 4:1-19.
- Chance B, Williams GR. Respiratory enzymes in oxidative phosphorylation. I. Kinetics of oxygen utilization. J Biol Chem. 1955;217(1):383-93.
- Estabrook RW. Mitochondrial respiratory control and polarographic measurement of ADP:O ration. Methods Enzymol. 1967;10:41-7.
- Buege J, Aust S. Microsomal lipid peroxidation. Methods Enzymol. 1978; LII :302-308.
- Huwiler M, Kohler H. Pseudo-catalytic degradation of hydrogen peroxide in the lactoperoxidase/H2O2/iodide system. Eur J Biochem 1984; 141: 69-74.
- Misra H, Fridovich I. The role of superoxide anion in the autoxidation of epinephrine and a simple assay superoxide dismutase. J Biol Chem. 1972; 247: 3170-75.
- Methods of Biochemical investigations. (Eds MI Prochorova). L: Izdatelstvo Leningrad universitet; 1982. [Russian].
- Rotruck J, Pope A, Ganther H, Swanson A. Selenium: biochemical role as a component of glutathione peroxidase. Science. 1973; 179: 588-90.
- Balashov Iu G. A fluorimetric micromethod for determining corticosteroids: a comparison with other methods. Fiziol Zh SSSR im IM Sechenova. 1990;76(2):280-83. [Russian].
- Zoccal D B, Bonagamba L, Antunes-Rodrigues J, Machado B H. Plasma corticosterone levels is elevated in rats submitted to chronic intermittent hypoxia. Autonomic Neurosci: Basic and Clin. 2007;134(1-2): 115–17.
- Wu Y, Du JZ. Effect of hypoxia on activity of hypothalamo-pituitary-adrenal-cortex axis in rat. Zhongguo Ying Yong Sheng Xue Za Zhi. 2001;17(4): 317-9.
- Zhi C, Ji-Zen D. Hypoxia effects on hypothalamic corticotropin-releasing hormone and anterior pituitary cAMP. Acta Pharm Sin. 1996;17(6): 489-92.
- Lukyanova LD.The modern problems of hypoxia.Vestnik RAMN 2000;9:3-12. [Russian].
- Maiti P, Shashi B S, Alpesh K S, Muthuraju S, Pratul K B, et al. Hypobaric hypoxia induces oxidative stress in rat brain. Neurochem Int. 2006; 49: 709-16.
- Pardo M, Tirosh O. Protective signaling effect of manganese superoxide dismutase in hypoxia-reoxygenation of hepatocytes. Free Rad Res. 2009; 43: 1225-39.
- Haddad J. Oxygen sensing mechanism and the regulation of redox-responsive transcription factors in development and pathophysiology. Respir Res. 2002; 3: 1-27.
- Kinnula V, Paakko P, Soini Y. Antioxidant enzymes and redox regulation thiol proteins in malignancies of human lung. FEBS Let. 2004; 569: 1-6.
- Sen C, Packer L. Antioxidant and redox regulation of gene transcription. FASEB J. 1996; 10: 709-20.
- Gonchar O, Mankovska I. Antioxidant system in adaptation to intermittent hypoxia. J Biol Sci. 2010; 10(6): 545-54.