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Effects of Exercise Intensity on Bcl-2, Bax, Caspase-3 Protein Level and DNA Fragmentation in Soleus and EDL Muscle on 60 wk SD Rats

운동강도의 차이가 60주령된 흰쥐의 Soleus와 EDL에서 Bcl-2, Bax, caspase-3와 DNA 절편화에 미치는 영향

  • Lee, Ji-Yung (Department of Physical Education, Dankook University) ;
  • Kim, Yong-An (Institute of Biotechnology, Chungnam Nat'l University)
  • 이지영 (단국대학교 사범대학 체육교육학과) ;
  • 김용안 (충남대학교 생물공학연구소)
  • Received : 2010.02.18
  • Accepted : 2010.04.22
  • Published : 2010.04.30

Abstract

The purpose of this study was to find out the Bcl-2 (B-cell leukemia/lymphoma-2), Bax, and caspase-3(cysteine-aspartic proteases-3) protein expression in soleus and EDL muscle according to treadmill exercise intensity in 60 week-old SD rats. The SD rats were randomly divided into four groups (n=10 in each group): control (CON), low intensity exercise (LE), moderate intensity exercise (ME), and high intensity exercise (HE). The exercise was given to the rats for 8 wk, 5 day/wk. The animals underwent treadmill exercise at intensities of 30 min at 8 m/min for the LE group, 15 min at 16 m/min for the ME group, and 9 min at 24 m/min for the HE group. The results were as follows: the expression of Bcl-2 protein was lowest in the HE group and the expression of Bax protein was highest in the HE group. The expression of caspase-3 (cleaved form) protein was observed in the HE group. For the different types of muscle fiber, Bcl-2 protein expression in the soleus muscle was decreased in all groups. Bax protein expression in the soleus muscle was increased in the HE group only. Bcl-2 protein expression in the EDL muscle was decreased in the HE group, and Bax protein expression in the EDL muscle was increased in the ME and HE groups. Consequently, the protein expression related to the aged rats shows a difference according to the intensity of exercise. In addition, caspase-3 protein expression appeared in the HE group; however, in all amounts of intensity, DNA fragmentation was not observed. Therefore, apoptosis on skeletal muscles of aged mice can be intervened with optimal exercise. On the other hand, high intensity exercise can potentially accelerate the apoptosis of muscle fiber in aged rats.

본 연구에서 60 주령된 쥐를 대상으로 8 주간 트레드밀 운동강도와 근섬유 유형에 따른 apoptosis 관련 인자를 연구하였다. 결론적으로 노후된 쥐의 apoptosis 관련 인자는 운동강도와 근육유형에 따라 차이가 났다. EDL이 Soleus보다 apoptosis인자의 변화가 운동강도에 의해 더 민감하게 나타났다. 또한 casepase-3의 발현은 고강도 운동에서 나타났으나 DNA 절편화는 나타나지 않았기에 실질적인 apoptosis는 발생되지 않았다. 이는 노후된 개체의 운동 수행방법으로 고강도 운동은 적합하지 않으며, 고강도 운동상태를 지속할 경우 근섬유의 apoptosis를 유도할 가능성이 있다는 것을 시사한다. 그러므로 추후 실험에서는 운동시간과 빈도에 의한 apoptosis의 time-course impact 관찰과 외재적 경로(caspase independent apoptosis)에서의 apoptosis 관련 단백질 변화에 대한 연구를 통하여 노화에 의한 근손실의 원인 규명이 요구된다.

Keywords

References

  1. Alway, S. E., H. Degens, G. Krishnamurthy, and A. Chaudhrai. 2003. Denervation stimulates apoptosis but not Id2 expression in hindimb muscles of aged rats. J. Gerontology Series A: Biol. Sci. Med. Sci. 58, 687-697. https://doi.org/10.1093/gerona/58.8.B687
  2. Armstrong, R., and R. Phelps. 1984. Muscle fiber type composition of ran hindlimp. Am. J. Anat. 171, 259-272. https://doi.org/10.1002/aja.1001710303
  3. Baumgartner, R. N., K. M. Koehler, D. Gallagher, L. Romero, S. B. Heymsfield, R. R. Ross, P. J. Garry, and R. D. Lindeman. 1998. Epidemiology of sarcopenioa among the elderly in New Mexico. Am. J. Epidemiol. 147, 755-763. https://doi.org/10.1093/oxfordjournals.aje.a009520
  4. Bedford, T. G., C. M. Tipton, N. C. Wilson, R. A. Oppliger, and C. V. Gisolfi. 1979. Maximum oxygen consumption of rats and its changes with various experimental procedures. J. Appl. Physiol. 47, 1278-1283.
  5. Chung, L. and Y. Ng. 2006. Age-related alterations in expression of apoptosis regulatory proteins and hear shock proteins in rat skeletal muscle. Biochim. Biophys. Acta. 1762, 103-109. https://doi.org/10.1016/j.bbadis.2005.08.003
  6. Concordet, J. P. and A. Ferry. 1993. Physiological programmed cell death in thymo cytes is induced by physical stress (exercise). Am. J. Physiol. 265, C626-C629.
  7. Danial, N. N., and S. J. Korsmeyer. 2004. Cell death: critical control points. Cell 116, 205-219. https://doi.org/10.1016/S0092-8674(04)00046-7
  8. Dirks, A. J. and C. Leeuwenburgh. 2006. Tumor necrosis factor $\alpha$ signaling in skeletal muscle: effects of age and caloric restriction. J. Nutri. Biochem. 17, 501-508. https://doi.org/10.1016/j.jnutbio.2005.11.002
  9. Dirks, A. J. and C. Leeuwenburgh. 2004. Aging and lifelong calorie restriction result in adaptation of skeletal muscle apoptosis repressor, apoptosis-inducing factor, X-linked inhibitor of apoptosis, caspase-3, and caspase-12. Free Radic. Biol. Med. 36, 27-39. https://doi.org/10.1016/j.freeradbiomed.2003.10.003
  10. Dirks, A. and C. Leeuwenburgh. 2002. Apoptosis in skeletal muscle with afina. Am. J. Physiol. Regul. Integr. Comp. Physiol. 282, 519-527.
  11. Dreyer, H. C. and E. Volpi. 2005. Role of protein and amino acids in the patho -physiology and treatment of sarcopenia. J. Am. Cell Nutr. 24, 140-145. https://doi.org/10.1080/07315724.2005.10719455
  12. Dupont-Versteegden, E. E. 2005. Apoptosis in muscle strophy: relevance to sarcopenia. Exp. Gerontol. 40, 473-418. https://doi.org/10.1016/j.exger.2005.04.003
  13. Ecochard, L., F. Lhenry, B. Sempore, and R. Favier. 2000. Skeletal muscle HSP72 level during endurance training: influence of peripheral arterial insufficiency. Pflugers Archv: European Journal of Physiology 440, 918-924. https://doi.org/10.1007/s004240000362
  14. Eva, M., A. Lisa, M. Cristina, B. Bert, L. Emanele, K. Masaaki, M. Daniel, R. Carlo, S. Stefano, and M. Sandri. 2009. Autophagy is required to maintain muscle mass. Cell Metabolism 10, 507-515. https://doi.org/10.1016/j.cmet.2009.10.008
  15. Hughes, V. A., W. R. Frontera, R. Roubenoff, W. F. Evans, and M. A. Singh. 2002. Longitudinal changes in body composition in older men and women: role of body weight and physical activity. Am. J. Clin. Nutr. 76, 473-481.
  16. Kannan, K. and S. K. Jain. 2000. Oxidative stress and apoptosis. Pathophysiology 7, 153-163. https://doi.org/10.1016/S0928-4680(00)00053-5
  17. Kocturk, S., B. M. Kayatekin, H. Rewmi, O. Acikgoz, C. Kaynak, and E. Ozer. 2008. The apoptotic response to strenuous exercise of the gastronemius and solues muscle fibers in rats. Eur. J. Appl. Physiol, 515-524.
  18. Leeuwenbergh, C., C. M. Gurley, B. A. Strotman, and E. E. Dupont-Versteegden. 2005. Age-related differences in apoptosis with disuse atrophy in Soleus muscle. Am. J. Physiol. integr. Comp. Physiol. 288, R1288-1296. https://doi.org/10.1152/ajpregu.00576.2004
  19. Phillips, T. and C. Leeuwenburgh. 2005. Muscle fiber specific apoptosis and TNF-alpha signaling in sarcopenia are attenuated by life-long calorie restriction. Br. J. Cancer 86, 1012-1016. https://doi.org/10.1038/sj.bjc.6600167
  20. Podhorska-Okolow, M., M. Sandri, S. Zampieri, B. Brun, K. Rossini, and U. Carraro. 1998. Apoptosis of myofibres and satellite cells: exercise-induced damage in skeletal muscle of the mouse. Neuropathol. Appl. Neurobiol. 24, 518-531. https://doi.org/10.1046/j.1365-2990.1998.00149.x
  21. Powers, S. K., D. Criswell, J. Lawler, L. L. Ji, D. R. Martin, A. Herb, G. Dudley. 1994. Influence of exercise and fiber type on antioxidant enzyme activity in rat skeletal muscle. Am. J. Physiol. 266, R375-R380.
  22. Rice, K. M. and E. R. Blough. 2006. Sarcopenia-selated apoptosis is regulated differently in fast- and slow-twitch muscles of the aging F344/N ${\times}$ BN rat model. Mechanism of Aging and Development 127, 670-679. https://doi.org/10.1016/j.mad.2006.03.005
  23. Rice, K. M. and E. R. Blough. 2006. Sarcopenia-related apoptosis is regulated differently in fast and slow twitch muscles of the aging F344/N ${\times}$ BN rat model. Mechanism of Ageing and Development 670-679.
  24. Sandri, M., U. Carraro, M. Ponhorska-Okolv, C. Rizzi., P. Arslan, D. Monti, and C. Franceschi. 1995. Apoptosis, DNA damage and ubiquitin expression in normal and mdx muscle fibers after exercise. FEBS. Lett. 373, 291-295. https://doi.org/10.1016/0014-5793(95)00908-R
  25. Sandri, M., M. Podhorska-Okolow, V. Geromel, C. Rizzi, P. I. Arslan, C. Franc-esche, and U. Carraro. 1997. Exercise indyces myonuclear ubiquitination and apoptosis in dystrophin-deficient muscle of mice. J. Neuropathol. Exp. Neurol.56, 45-57. https://doi.org/10.1097/00005072-199701000-00005
  26. Siu, P. M., R. W. Bruner, J. K. Martyn, and S. E. Alway. 2004. Apoptotic adaptations from exercise training in skeletal and cardiac muscles. FASEB. J. 18, 1150-1152.
  27. Siu, P. M., E. E. Pistilli, M. J. Ryan, and S. E. Alway. 2005. Aging sustains the hypertrophy-associated elevation of apoptotic suppressor X-linked inhibitor of apoptotic protein (XIAP) in skeletal muscle during unloading. H. Gerontol. A Biol. Sci. Med. Sci. 60, 976-983. https://doi.org/10.1093/gerona/60.8.976
  28. Siu, P. M. and S. E. Alway. 2006. Aging alters the reduction of pro-apoptotic signaling to loading-induced hyper-trophy. Exp. Gerontol. 41, 175-188. https://doi.org/10.1016/j.exger.2005.11.004
  29. Smith, J. 1996. Muscle growth factors, ubiqutin and apoptosis in dystrophic muscle: apoptosis decline with age in the mdx mouse. Basic. Appl. Myol. 6, 279-284.
  30. Song, W., H. B. Kwak, and J. M. Lawler. 2006. Exercise training attenuates age-induced changes in apoptotic signaling in rat skeletal muscle. Antioxid. Redox Signal. 8, 517-528. https://doi.org/10.1089/ars.2006.8.517
  31. Szulc, P., F. Duboeuf, F. Marchand, and P. D. Delmas. 2004. Hormonal and lifestyle determinants of appendicular skeletal muscle mass in men; the MINOS study. Am. J. Clin. Nutr. 80, 496-503.
  32. Thompson, H. S., P. M. Clarkson, and S. F. Scordilis. 2002. The repeated bout effect and heat shock proteins: Intramuscular HSP27 and HSP70 expression following two bouts of eccentric exercise in human. Acta Physiolgica Scandinavita 174, 47-56. https://doi.org/10.1046/j.1365-201x.2002.00922.x
  33. Tidball, J. G., D. E. Albrecht, B. E. Locknsgard, and M. J. Spencer. 1995. Apoptosis precedes necrosis ofdystrophin-deficient muscle. J. Cell Sci. 108, 2197-2204.
  34. Vandervoort, A. A. 2002. Aging of the human neuromuscular system. Muscle Nerve. 25, 17-25. https://doi.org/10.1002/mus.1215
  35. Wyille, A. H., J. F. Kerrm, and A. R. Curie. 1980. Cell death: the significance of apoptosis. Int. Rev. Cytol. 68, 251-305. https://doi.org/10.1016/S0074-7696(08)62312-8