Skip to main content
SpringerLink
Log in

Rabbit annulus fibrosus cell apoptosis induced by mechanical overload via a mitochondrial apoptotic pathway

  • Published:
Journal of Huazhong University of Science and Technology [Medical Sciences] Aims and scope Submit manuscript

Summary

In order to investigate the apoptotic pathway of rabbit annulus fibrosus (AF) cells induced by mechanical overload, an experimental air-pressure model was established in this study to pressurize the rabbit AF cells in vitro. Cells were randomly divided into five groups in which the cells were exposed to a continuous pressure of 1.1 MPa for different lengths of time (0, 5, 12, 24 and 36 h). The cell proliferation and apoptosis were detected by cell counting kit-8 (CCK-8) assay and flow cytometry; the alterations in mitochondrial membrane potential were measured by fluorescence microscopy and fluorescence spectrophotometer; the activities of caspase-8 and 9 were determined by spectrophotometry. The results showed that after the cells were subjected to the pressure for 24 or 36 h, the cell proliferation was inhibited; the ratio of cell apoptosis was increased; the mitochondrial membrane potential was decreased; the activity of caspase-9 was enhanced; no activity changes were observed in caspase-8. The results suggested that treatment with a pressure of 1.1 MPa for more than 24 h can lead to the proliferation inhibition and the apoptosis of rabbit AF cells in vitro, and the mitochondrial-dependent pathway is implicated in the pressure-induced AF cell apoptosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Kasra M, Merryman WD, Loveless KN, et al. Frequency response of pig intervertebral disc cells subjected to dynamic hydrostatic pressure. J Orthop Res, 2006,24(10):1967–1973

    Article  PubMed  Google Scholar 

  2. Ali R, Le MCL, Richardson SM, et al. Connective tissue growth factor expression in human intervertebral disc: implications for angiogenesis in intervertebral disc degeneration. Biotech Histochem, 2008, 83(5):239–245

    Article  CAS  PubMed  Google Scholar 

  3. Hoogendoorn R, Doulabi BZ, Huang CL, et al. Molecular changes in the degenerated goat intervertebral disc. Spine (Phila Pa 1976), 2008, 33(16):1714–1721

    Google Scholar 

  4. Le MCL, Freemont AJ, Hoyland JA. Accelerated cellular senescence in degenerate intervertebral discs: a possible role in the pathogenesis of intervertebral disc degeneration. Arthritis Res Ther, 2007,9(3):R45

    Article  Google Scholar 

  5. Junger S, Gantenbein-Ritter B, Lezuo P, et al. Effect of limited nutrition on in situ intervertebral disc cells under simulated-physiological loading. Spine (Phila Pa 1976), 2009,34(12):1264–1271

    Google Scholar 

  6. Rajasekaran S, Venkatadass K, Naresh BJ, et al. Pharmacological enhancement of disc diffusion and differentiation of healthy, ageing and degenerated discs: Results from in-vivo serial post-contrast MRI studies in 365 human lumbar discs. Eur Spine J, 2008,17(5):626–643

    Article  CAS  PubMed  Google Scholar 

  7. Tschoeke SK, Hellmuth M, Hostmann A, et al. Apoptosis of human intervertebral discs after trauma compares to degenerated discs involving both receptor-mediated and mitochondrial-dependent pathways. J Orthop Res, 2008, 26(7):999–1006

    Article  CAS  PubMed  Google Scholar 

  8. Wei A, Brisby H, Chung SA, et al. Bone morphogenetic protein-7 protects human intervertebral disc cells in vitro from apoptosis. Spine J, 2008,8(3):466–474

    Article  PubMed  Google Scholar 

  9. Zhao CQ, Jiang LS, Dai LY. Programmed cell death in intervertebral disc degeneration. Apoptosis, 2006,11(12): 2079–2088

    Article  PubMed  Google Scholar 

  10. Jones P, Gardner L, Menage J, et al. Intervertebral disc cells as competent phagocytes in vitro: implications for cell death in disc degeneration. Arthritis Res Ther, 2008,10(4):R86

    Article  PubMed  Google Scholar 

  11. Lotz JC, Chin JR. Intervertebral disc cell death is dependent on the magnitude and duration of spinal loading. Spine (Phila Pa 1976), 2000,25(12):1477–1483

    CAS  Google Scholar 

  12. Wuertz K, Urban JP, Klasen J, et al. Influence of extracellular osmolarity and mechanical stimulation on gene expression of intervertebral disc cells. J Orthop Res, 2007,25(11):1513–1522

    Article  CAS  PubMed  Google Scholar 

  13. Lee CR, Iatridis JC, Poveda L, et al. In vitro organ culture of the bovine intervertebral disc: effects of vertebral endplate and potential for mechanobiology studies. Spine (Phila Pa 1976), 2006,31(5):515–522

    Google Scholar 

  14. Kasra M, Goel V, Martin J, et al. Effect of dynamic hydrostatic pressure on rabbit intervertebral disc cells. J Orthop Res, 2003,21(4):597–603

    Article  PubMed  Google Scholar 

  15. Yu WR, Liu T, Fehlings TK, et al. Involvement of mitochondrial signaling pathways in the mechanism of Fas-mediated apoptosis after spinal cord injury. Eur J Neurosci, 2009,29(1): 114–131

    Article  PubMed  Google Scholar 

  16. Rannou F, Lee TS, Zhou RH, et al. Intervertebral disc degeneration: the role of the mitochondrial pathway in annulus fibrosus cell apoptosis induced by overload. Am J Pathol, 2004,164(3):915–924

    CAS  PubMed  Google Scholar 

  17. Park JB, Park IC, Park SJ, et al. Anti-apoptotic effects of caspase inhibitors on rat intervertebral disc cells. J Bone Joint Surg Am, 2006,88(4):771–779

    Article  PubMed  Google Scholar 

  18. Ozaki T, Iizuka K, Suzuki M, et al. Threshold-dependent DNA synthesis by pure pressure in human aortic smooth muscle cells: Gialpha-dependent and -independent pathways. Biochem Biophys Res Commun, 1999,256(1): 212–217

    Article  CAS  PubMed  Google Scholar 

  19. Hutton WC, Elmer WA, Boden SD, et al. The effect of hydrostatic pressure on intervertebral disc metabolism. Spine (Phila Pa 1976), 1999,24(15):1507–1515

    CAS  Google Scholar 

  20. Kroeber M, Unglaub F, Guehring T, et al. Effects of controlled dynamic disc distraction on degenerated intervertebral discs: an in vivo study on the rabbit lumbar spine model. Spine (Phila Pa 1976), 2005,30(2):181–187

    Google Scholar 

  21. Unglaub F, Guehring T, Omlor G, et al. Controlled distraction as a therapeutic option in moderate degeneration of the intervertebral disc — an in vivo study in the rabbit-spine model. Z Orthop Ihre Grenzgeb, 2006,144(1): 68–73

    Article  CAS  PubMed  Google Scholar 

  22. Liu GZ, Ishihara H, Osada R, et al. Nitric oxide mediates the change of proteoglycan synthesis in the human lumbar intervertebral disc in response to hydrostatic pressure. Spine (Phila Pa 1976), 2001,26(2):134–141

    CAS  Google Scholar 

  23. Yu JS, Qiu GX, Yang BL, et al. The difference and its significance of actin in inverterbral disc cells under cyclic hydrostatic pressure in vitro. Chin J Orthopaedics, 2005, (09):52–56

  24. Ariga K, Yonenobu K, Nakase T, et al. Mechanical stress-induced apoptosis of endplate chondrocytes in organ-cultured mouse intervertebral discs: an ex vivo study. Spine (Phila Pa 1976), 2003,28(14):1528–1533

    Google Scholar 

  25. Wilke HJ, Neef P, Caimi M, et al. New in vivo measurements of pressures in the intervertebral disc in daily life. Spine (Phila Pa 1976), 1999,24(8):755–762

    CAS  Google Scholar 

  26. Hishikawa K, Oemar BS, Nakaki T. Static pressure regulates connective tissue growth factor expression in human mesangial cells. J Biol Chem, 2001,276(20):16797–16803

    Article  CAS  PubMed  Google Scholar 

  27. Le MCL, Frain J, Fotheringham AP, et al. Human cells derived from degenerate intervertebral discs respond differently to those derived from non-degenerate intervertebral discs following application of dynamic hydrostatic pressure. Biorheology, 2008,45(5):563–575

    Google Scholar 

  28. Scoltock AB, Cidlowski JA. Activation of intrinsic and extrinsic pathways in apoptotic signaling during UV-C-induced death of Jurkat cells: the role of caspase inhibition. Exp Cell Res, 2004,297(1):212–223

    Article  CAS  PubMed  Google Scholar 

  29. Schuler M, Bossy-Wetzel E, Goldstein JC, et al. p53 induces apoptosis by caspase activation through mitochondrial cytochrome C release. J Biol Chem, 2000,275(10): 7337–7342

    Article  CAS  PubMed  Google Scholar 

  30. Mazumder S, Plesca D, Almasan A. Caspase-3 activation is a critical determinant of genotoxic stress-induced apoptosis. Methods Mol Biol, 2008,414:13–21

    Article  CAS  PubMed  Google Scholar 

  31. Rudner J, Lepple-Wienhues A, Budach W, et al. Wild-type, mitochondrial and ER-restricted Bcl-2 inhibit DNA damage-induced apoptosis but do not affect death receptor-induced apoptosis. J Cell Sci, 2001,114(Pt 23): 4161–4172

    CAS  PubMed  Google Scholar 

  32. Andrews HE, Nichols PP, Bates D, et al. Mitochondrial dysfunction plays a key role in progressive axonal loss in multiple sclerosis. Med Hypotheses, 2005,64(4):669–677

    Article  CAS  PubMed  Google Scholar 

  33. van RBJ, Verhoeven AJ, Kuijpers TW. Mitochondria in neutrophil apoptosis. Int J Hematol, 2006,84(3):199–204

    Article  Google Scholar 

  34. Eeva J, Nuutinen U, Ropponen A, et al. Feedback regulation of mitochondria by caspase-9 in the B cell receptor-mediated apoptosis. Scand J Immunol, 2009,70(6): 574–583

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China

    Mao Xie  (谢 卯), Shuhua Yang  (杨述华), Hein Latt Win, Liming Xiong  (熊蠡茗), Jijun Huang  (黄吉军) & Jianguo Zhou  (周建国)

Authors
  1. Mao Xie  (谢 卯)
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shuhua Yang  (杨述华)
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hein Latt Win
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Liming Xiong  (熊蠡茗)
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jijun Huang  (黄吉军)
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jianguo Zhou  (周建国)
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Liming Xiong  (熊蠡茗).

Additional information

This project was supported by a grant from National Natural Sciences Foundation of China (No. 30700841).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Xie, M., Yang, S., Win, H.L. et al. Rabbit annulus fibrosus cell apoptosis induced by mechanical overload via a mitochondrial apoptotic pathway. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 30, 379–384 (2010). https://doi.org/10.1007/s11596-010-0361-4

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11596-010-0361-4

Key words

Search

Navigation