EXPRESSION OF MITOCHONDRIAL UNCOUPLING PROTEIN 3 AND THE SENSITIVITY OF MITOCHONDRIAL PERMEABILITY TRANSITION PORE OPENING TO CA2+ IN OLD RAT HEART UNDER ACTIVATION OF BIOSYNTHESIS OF COENZYME Q
N. A. Strutynska, S.V. Timoshchuk, G.L.Vavilova, A.V. Kotsuruba, V.F.Sagach
O.O. Bogomoletz Institute of Physiology NationalAcademy Sciences of Ukraine, Kyiv, Ukraine
The expression of mitochondrial uncoupling protein 3 (UCP3), as well as the sensitivity of mitochondrial permeability transition pore opening (MPTP) to Ca2+ (10-4 mol/l) in old rat heart under activation in vivo of ubiquinone synthesis - coenzyme Q, (CoQ) via administration of the precursors (4-hydroxybenzoic acid, aminoacid methionine and modulator vitamin E) were studied. It was shown that the expression level of UCP3 decreased by 63% in old rats compared to adult rats and this was accompanied by an increased sensitivity of the MPT to calcium. Under activation of endogenous synthesis of CoQ it was observed almost complete restoration of UCP3 expression in old rat heart and a decrease in the sensitivity of the MPTP opening to Ca2+. In mitochondria from old rat hearts we noted an increased content of the superoxide (O2) and hydroxyl (яOH) radicals and of the stable metabolite of active oxygen species hydrogen peroxide (HO), as compared to those in adult animals. Following activation of endogenous synthesis of CoQ in old rat heart mitochondria it was observed a decreased content of H2O2, andthe tendency for decreasing the levels of the radicals яO2 and яOH. The results obtained allowed to conclude that the CoQ-dependent restoration of the UCP3 levels in old rat heart and antioxidant/cardioprotective effects of CoQ related to the MPTP opening inhibition can reduce the oxidative stress and thus prevent the manifestation of mitochondrial dysfunction in aging heart. We suggest that UCP3 is not involved in the increase of the passive H+-conductance through the inner mitochondrial membrane in the aging heart, and that CoQ as a factor of respiratory chain could be an important endogenous regulator of the uncoupling proteins, in particular UCP3, in the heart. Key words: UCP 3 protein, mitochondrial permeability transition pore, coenzyme Q, active oxygen radicals, aging, rat.
mitochondrial uncoupling protein 3, mitochondrial permeability transition pore
- Sagach VF, Vavilova GL, Rudyk OV etc. Inhibition of mitochondrial pores as one of the mechanisms of cardioprotective action of coenzyme Q . Fiziol Zh. 2007. 53, N 4. p 35-42.
- Sagach VF, Vavilova GL, Strutynska NA, Rudyk OV Aging increases the sensitivity to inducers of mitochondrial pores in the heart of rats . Fiziol Zh. 2004. 50, N 2. p 49-63.
- Sagach VF, Rudyk OV, Vavilova GL etc. Melatonin restores ischemic tolerance and reduces the sensitivity of the opening of mitochondrial pores in the heart of old rats . Fiziol Zh. 2006. 52, N 3. p 3-14.
- Timoshchuk SV, Vavilova GL, Strutynska NA etc. Influence of ubiquinone biosynthesis precursors in vivo on the sensitivity of mitochondrial pore opening in the heart of old rats . Fiziol Zh. 2008. 54, N 3. P.3-9.
- Amaral S., Mota P., Rodrigues A.S. et al. Testicular aging involves mitochondrial dysfunction as well as an increase in UCP2 levels and proton leak . FEBS Lett. 2008. 582. P. 4191-4196.
- Barazzoni R., Nair K.S. Changes in uncoupling pro-tein-2 and 3 expression in aging rat skeletal muscle, liver, and heart . Amer. J. Phys. Endocrinol. Met. 2001. 280, N 3. P.413-419.
- Berzaire V., Seifert E. L., Harper M-E. Uncoupling protein-3: clues in an ongoing mitochondrial mystery . . J. FASEB. 2007. 21. P.312-324.
- Bo H., Jiang N., Ma G. et al. Regulation of mitochondrial uncoupling respiration during exercise in rat heart: role of reactive oxygen species (ROS) and uncoupling protein 2 . Free Radic. Biol. and Med. 2008. 44, N 7. P.1373-1381.
- Boss O., Hagen T., Lowell B.B. Uncoupling proteins 2 and 3. Potential regulators of mitochondrial energy metabolism . Diabetes. 2000. 49. P.143-156.
- Cannon B., Shabalina I.G., Kramarova T.V. et al. Uncoupling proteins: A role in protection against reactive oxygen species or not? . Biochim. and Biophys. Acta. 2006. 1757. P. 449-458.
- Echtay K.S., Murphy M.P., Smith R.A. J. et al. Superoxide activates nucleotide-sensitive mitochondrial proton transport through the uncoupling proteins UCP1, UCP2, and UCP3 . Nature. 2002. 415. P. 1482-1486.
- Essop M. F., Razeghi P., McLeod C. et al. Hypoxia-induced decrease of UCP3 gene expression in rat heart parallels metabolic gene switching but fails to affect mitochondrial respiratory coupling . Biochim. and Biophys. Res. Communs. 2004. 314. P. 561-564.
- Fernstrom M., Tonkonogi M., Sahlin K. Effects of acute and chronic endurance exercise on mitochondrial uncoupling in human skeletal muscle . J. Physiol. 2003. 554,N 3. P.755-763.
- Fontaine E., Ichas F., Bernardi P. A ubiquinone-bind-ing site regulates the mitochondrial permeability transition pore . J. Biol. Chem. 1998. 273, N 40. P.25734-25740.
- Graier W.F., Trenker M., Malli R. Mitochondrial Ca2+, the secret behind the function of uncoupling proteins 2 and 3? . Cell Calcium. 2008. 44. P. 36-50.
- Harman D. The aging process . Proc. Natl. Acad Sci. USA 1981. 78. P.7124-7128.
- He L., Lemasters J.J. Regulated and unregulated mitochondrial permeability transition pores: a new paradigm of pore structure and function? . FEBS Lett. 2002. 512. P.1-7.
- Hirabara S.M., Silveira L.R., Abdulkader F.R.M. et al. Role of fatty acids in the transition from anaerobic to aerobic metabolism in skeletal muscle during exercise . Cell Biochem. Funct. 2006. 24. P.475-481.
- Juhaszova M., Wang S., Zorov D.B. et al. The identity and regulation of the mitochondrial permeability transition pore: where the known meets the unknown . Ann. N-Y Acad. Sci. 2008. 1123. P.197-212.
- Kerner J., Turkaly P.J., Minkler P.E.,. Hoppel C.L. Aging skeletal muscle mitochondria in the rat: decreased uncoupling protein-3 content . Amer. J. Physiol. Endocrinol. Metab. 2001. 281. P. 1054-1062.
- Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4 . Nature. 1970. 227. P. 680-685.
- Lesnefsky E.J., Hoppel C.L. Ischemia-reperfusion injury in the aged heart: role of the mitochondria . Arch. Biochem. and Biophys. 2003. 420, N 2. P.287-297.
- Marcinek D.J., Schenkman K.A., Ciesielski W.A. et al. Reduced mitochondrial coupling in vivo alters cellular energetics in aged mouse skeletal muscle . J. Physiol. 2005. 569,N 2 P.467-473.
- McLeod C.J., Aziz A., Hoyt R. F. et al. Uncoupling proteins 2 and 3 function in concert to augment tolerance to cardiac ischemia . J. Biol. Chem. 2005. 280, N 39. P. 33470-33476.
- Murray A.J., Cole M.A., Lygate C.A. et al. Increased mitochondrial uncoupling proteins, respiratory uncou-pling and decreased efficiency in the chronically infarcted rat heart . J. Mol. and Cell. Cardiol. 2008. 44. P. 694-700.
- Nabben M., Hoeks J. Mitochondrial uncoupling protein 3 and its role in cardiac- and skeletal muscle metabolism . Physiol. and Behavior. 2008. 94. P. 259-269.
- Nedergaard J., Cannon B. The ‘novel’ ‘uncoupling’ proteins UCP2 and UCP3: what do they really do? Pros and cons for suggested functions . J. Exp. Physiol. 2003. 88, N 1. P. 65-84.
- O’Rourke B. Mitochondrial ion channels . Ann. Rev. Physiol. 2007. 69. P.19-49.
- Pepe S. Mitochondrial function in ischaemia and reperfusion of the ageing heart . Clin. and Exp. Pharmacol. and Physiol. 2001. 27, N 9. P.745-750.
- Razeghi P., Young M.E., Abbasi S., Taegtmeyer H. Hypoxia in vivo decreases peroxisome proliferator-activated receptor alpha-regulated gene expression in rat heart . Biochem. and Biophys. Res. Communs. 2001. 287. P. 5-10.
- Ricquier D., Bouillaud F. Mitochondrial uncoupling proteins: from mitochondria to the regulation of energy balance .J. Physiol. 2000. 529. P.3-10.
- Sohal R.S., Forster M.J. Coenzyme Q, oxidative stress and aging . Mitochondrion. 2007. 7S P.103-111.
- Turunen M., Olsson J., Dallner G. Metabolism and function coenzyme Q . Biochim. and Biophys. Acta 2004. 1660. P. 171-199.