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Role of mitochondrial pathway in compression-induced apoptosis of nucleus pulposus cells

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Abstract

Various mechanical stresses can induce apoptosis of nucleus pulposus (NP) cells and intervertebral disc (IVD) degeneration in vivo, but the underlying molecular mechanism by which the number of NP cells is decreased in degenerated IVD is still not elucidated. The purpose of this study was to investigate whether the mitochondrial pathway is involved in compression-induced apoptosis of rabbit NP cells. The compression apparatus was used to investigate the effect of the compression in this process at one magnitude (1.0 MPa) for 6, 12, 18, 24 and 36 h. Cell viability was measured by cell counting kit-8. Apoptosis rate was analyzed by flow cytometry and the morphologic changes in apoptosis cells were observed by the phase-contrast microscopy and Hoechst 33258 staining. The apoptosis-related gene and protein synthesis, such as Bax, Bcl-2 and Caspase-3, was analyzed by real-time polymerase chain reaction and Western-blot, respectively. Mitochondrial function was evaluated by analyzing the mitochondrial permeability transition pore (MPTP), as well as reactive oxygen species (ROS) and mitochondrial membrane potential (MMP). The results indicated that compression at the magnitude of all time points induced apoptosis of rabbit NP cells in a time-dependent manner, and the cell viability was reduced significantly. Furthermore, the compression at this level profoundly suppressed the functions of the mitochondria such as the opening of MPTP, the excessive production of ROS and the decreased MMP. Our findings suggest that the compression-induced IVD degeneration is mediated, at least in part, via the mitochondrial apoptotic pathway in NP cells.

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Acknowledgments

This work was supported by a grant from National Natural Sciences Foundation of China (No. 30700841, 81171149). Special thanks go to the members of the IVD degeneration research team at the Institute of Orthopedic Surgery of Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, the People’s Republic of China for their help.

Conflict of interest

Foundation funds were received in support of this work. No benefits in any form have been or will be received from a commercial party related directly or indirectly to the subject of this manuscript. The authors declare that they have no conflict of interest.

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Authors and Affiliations

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

    Fan Ding, Zeng-Wu Shao, Shu-Hua Yang, Qiang Wu, Fei Gao & Li-Ming Xiong

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  1. Fan Ding
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  2. Zeng-Wu Shao
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  3. Shu-Hua Yang
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Correspondence to Li-Ming Xiong.

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Fan Ding and Zeng-Wu Shao contributed equally to this work.

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Ding, F., Shao, ZW., Yang, SH. et al. Role of mitochondrial pathway in compression-induced apoptosis of nucleus pulposus cells. Apoptosis 17, 579–590 (2012). https://doi.org/10.1007/s10495-012-0708-3

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