DOI QR코드

DOI QR Code

8주간 운동이 생쥐의 gastrocnemius에서 Bcl-2, Bax, caspase-8, caspase-3와 HSP70에 미치는 영향

Effects of 8-week Exercise on Bcl-2, Bax, Caspase-8, Caspase-3 and HSP70 in Mouse Gastrocnemius Muscle

  • 김기범 (한신대학교 특수체육학과) ;
  • 김용안 (충남대학교 생물공학연구소) ;
  • 박정준 (부산대학교 스포츠과학부)
  • Kim, Ki-Bum (Department of Adapted Physical Education, Hanshin University) ;
  • Kim, Yong-An (Institute of Biotechnology, Chungnam Nattional University) ;
  • Park, Jung-Jun (Division of Sports Science, Pusan Nattional University)
  • 투고 : 2010.08.09
  • 심사 : 2010.09.10
  • 발행 : 2010.09.30

초록

이 연구는 실험용 쥐를 대상으로 운동에 따라 골격근에서 일어나는 세포고사 경로 중 내인성 경로와 외인성경로에 의해 이루어지는 세포고사의 신호기전을 확인하여 운동이 세포고사에 어떠한 영향을 미치는지에 대한 구명하는데 있다. 이를 위해 ICR계 수컷 흰쥐 20마리를 일주일의 적응기간을 거쳐 통제집단(CON: n=10)과 운동집단(EX: n=10)으로 배정하였다. 운동은 8주간 주 5회 실시하였고, 트레드밀 속도 16.4 m/min와 경사도 4%로 설정하여 40분간 지속적인 운동을 실시하였다. 세포고사의 신호 경로 중 내인성 경로에 대한 검증 결과 Bcl-2, Bax 단백질, 그리고 Bcl-2/ Bax ratio는 그룹간 통계적 유의성은 나타나지 않았다. 반면, 세포고사의 경로 중 외인성 경로에 대한 검증 결과 caspase-8 단백질의 발현은 운동집단이 통제집단보다 유의하게 낮은 것으로 나타났으며(p<0.05), 세포고사 억제 단백질인 HSP70 단백질 발현은 운동집단이 통제집단보다 높게 나타났다. 더욱이, 세포고사 최종인자인 caspase-3의 활성화는 이루어 지지 않은 것으로 관찰되었다. 따라서 세포사멸의 신호경로 중 내인성 경로에 작용하는 Bcl-2와 Bax보다는 외인성 경로인 caspase-8과 HSP70의 영향으로 caspase-3가 분할되지 못하여 세포고사가 일어나지 않은 것으로 사료된다.

The aim of this study was to investigate the effects of exercise on intrinsic and extrinsic apoptosis signaling pathways in skeletal muscle. ICR-type white male mice were divided into a control group (CON: n=10) and an exercise training group (EX: n=10) after a 1 week adaptation period. EX performed treadmill running at 16.4 m/min with a 4% incline, 40 min/day and 5 days/week for 8 weeks. Cervical dislocation was performed at 48 hours after the last bout of exercise, after which gastrocnemius skeletal muscles were immediately collected. The results of verifying the intrinsic apoptosis pathway showed that there were no significant differences in Bcl-2, Bax, or the ratio of Bax/Bcl-2 proteins between EX and CON. On the other hand, the results of verifying the extrinsic apoptosis pathway showed that caspase-8 proteins were significantly lower in EX than in CON (p<0.05). Apoptosis suppressing protein HSP70 was higher in EX than in CON. In addition, caspase-3, which is the final factor for apoptosis, was not activated. These results indicate that apoptosis did not develop since caspase-3 is non-cleaved by the effects of caspase-8 and HSP70 extrinsic pathways rather than Bcl-2 and Bax intrinsic pathways among signal pathways for apoptosis.

키워드

참고문헌

  1. Cande, C., I. Cohen., E. Daugas., L. Ravagnan., N. Larochette., N. Zamzami., and G. Kroemer. 2002. Apoptosis-inducing factor (AIF): a novel caspase-independent death effector released from mitochondria. Biochimie. 84, 215-222. https://doi.org/10.1016/S0300-9084(02)01374-3
  2. Chae, C. H. and H. T. Kim. 2009. Forced, moderate-intensity treadmill exercise suppresses apoptosis by increasing the level of NGF and stimulating phosphatidylinositol 3-kinase signaling in the hippocampus of induced aging rats. Neurochem. Int. 55, 208-213. https://doi.org/10.1016/j.neuint.2009.02.024
  3. Cory, S. and J. Adams. 2002. The Bcl-2 Family: regulators of the cellular life-or-death switch. Nat. Rev. Cancer 2, 647-656. https://doi.org/10.1038/nrc883
  4. Droge, W. 2002. Free radicals in the physiological control of cell function. Physio. Rev. 82, 47-95.
  5. Emidio, E. P., R. J. Janna., and E. A. Stephen. 2006. Death receptor- associated pro-apoptotic signaling in aged skeletal muscle. Apoptosis 11, 2115-2126. https://doi.org/10.1007/s10495-006-0194-6
  6. Ghosh, S., M. Khazaei., F. Moien-Afshari., L. S. Ang., D. J. Granville., C. B. Verchere., S. R. Dunn., P. McCue., A. Mizisin., K. Sharma, and I. Laher. 2009. Moderate exercise attenuates caspase-3 activity, oxidative stress, and inhibits progression of diabetic renal disease in db/db mice. Am. J. Physiol. Renal. Physiol. 296, 700-708. https://doi.org/10.1152/ajprenal.90548.2008
  7. Green, D. R. and J. Reed. 1998. Mitochondria and apoptosis. Science 281, 1309-1312. https://doi.org/10.1126/science.281.5381.1309
  8. Gross, A., J. M. Mcdonnell, and S. J. Korsmeyer. 1999. Bcl-2 family members and the mitochondria in apoptosis. Gene Dev. 13, 1899-1911. https://doi.org/10.1101/gad.13.15.1899
  9. Gruter, M. G. 2000. Caspases: Key players in programmed cell death. Curr. Opinstruct Biol. 10, 649-655.
  10. Harkins, M. S. 2009. Exercise regulates heat shock proteins and nitric oxide. Exerc. Sport Sci. Rev. 37, 73-77. https://doi.org/10.1097/JES.0b013e31819c2e7a
  11. Hengartner, M. O. 2000. The biochemistry of apoptosis. Nature 407, 770-776. https://doi.org/10.1038/35037710
  12. Ji, L. L. 2002. Exercise-induced modulation of antioxidant defense. Ann. N. Y. Acad. Sci. 959, 82-92.
  13. 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
  14. Kocturk, S., B. M. Kayatekin, H. Resmi, O. Acikgoz, C. Kaynak, and E. Ozer. 2008. The apoptotic response to strenuous exercise of the gastrocnemius and solues muscle fibers in rats. Eu. J. Appl. Physiol. 102, 515-524. https://doi.org/10.1007/s00421-007-0612-7
  15. Koctüvrk, S., B. M. Kayatekin, H. Rewmi, O. Acrivkgorz, C. Kaynak, and E. Övzer. 2008. The apoptotic response to strenuous exercise of the gastronemius and solues muscle fibers in rats. Eur. J. Appl. Physiol. 8, 515-524.
  16. Kroemer, G. 2003. Mitochondrial control of apoptosis: an introduction. Biochem. Biophys. Res. Commun. 304, 433-435. https://doi.org/10.1016/S0006-291X(03)00614-4
  17. Lee, J. S., K. C. Park, D. S. Kim, H. O. Choi, K. H. Kim, J. H. Chung, H. C. Eun, and J. S. Seo. 2000. Overexpression of HSP70 prevents ultraviolet B-induced apoptosis of a human melanoma cell line. Arch. Dermatol. Res. 292, 482-487. https://doi.org/10.1007/s004030000173
  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. Lim, M. L., M. G. Lum, T. M. Hansen, X. Roucou, and P. Nagley. 2002. On the release of cytochrome c from mitochondria during cell death signaling. J. Biomed. Sci. 9, 288-506. https://doi.org/10.1007/BF02256075
  20. Martinadle, J. L. and N. J. Holbrook. 2002. Cellular response to oxidative stress: signaling for suicide and survival. J. Cell Physiol. 192, 1-15. https://doi.org/10.1002/jcp.10119
  21. Morimoto, R. I. 1998. Regulation of the heat shock transcriptional response: cross talk between a family of heat shock factors, molecular chaperones, and negative regulators. Genes Dev. 12, 3788-3796. https://doi.org/10.1101/gad.12.24.3788
  22. Phaneuf, S. and C. Leeuwenburgh. 2001. Apoptosis and exercise. Med. Sci. Sports Exerc. 33, 393-396. https://doi.org/10.1097/00005768-200103000-00010
  23. Phillips, S. 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
  24. 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
  25. Powers, S. K., D. Criswell, J. Lawler, L. L. Ji, D. R. Martin, A. Herb, and G. Dudley. 1994. Influence of exercise and fiber type on antioxidant enzyme activity in rat skeletal muscle. Am. J. Physiol. 266, R375-R380.
  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, 1150-2.
  27. Tang, D. and V. J. Kidd. 2000. Caspase-8 activation and bid cleavage contribute to MCF7 cellular execution in a caspase- 3-dependent manner during staurosporine-mediated apoptosis. J. Biol. Chem. 275, 9303-9307. https://doi.org/10.1074/jbc.275.13.9303
  28. Thambirajah, A. A., K. Sleigh, H. G. Stiver, and A. W. Chow. 2008. Differential heat shock protein responses to strenuous standardized exercise in chronic fatigue syndrome patients and matched healthy controls. Clin. Invest Med. 31, 319-327.
  29. 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
  30. Tidball, J. G., D. E. Albrecht, B. E. Locknsgard, and M. J. Spencer. 1995. Apoptosis precedes necrosis of dystrophin- deficient muscle. J. Cell Sci. 108, 2197-2204.
  31. Wang, J. S. and Y. Huang. 2005. Effects of exercise intensity on lymphocyte apoptosis induced by oxidative stress in men. Eur. J. Appl. Physiol. 95, 290-297. https://doi.org/10.1007/s00421-005-0005-8
  32. 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
  33. Ye, H., C. Cande., N. C. Stephanou., S. Jiang., S. Gurbuxani., N. Larochette., E. Daugas., C. Garrido., G. Kroemer, and H. Wu. 2002. DNA binding is required for the apoptogenic action of apoptosis inducing factor. Nat. Struct Biol. 9, 680-684. https://doi.org/10.1038/nsb836

피인용 문헌

  1. The Autophagic Response to Exercise Training of the Skeletal Muscle Fibers in Young and Old Mice vol.21, pp.3, 2011, https://doi.org/10.5352/JLS.2011.21.3.400
  2. Effects of Liriopis Tuber on 4-HNE-induced Apoptosis in PC-12 Cells vol.28, pp.2, 2013, https://doi.org/10.6116/kjh.2013.28.2.33