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Beta1 integrin inhibits apoptosis induced by cyclic stretch in annulus fibrosus cells via ERK1/2 MAPK pathway

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Abstract

Low back pain is associated with intervertebral disc degeneration (IVDD) due to cellular loss through apoptosis. Mechanical factors play an important role in maintaining the survival of the annulus fibrosus (AF) cells and the deposition of extracellular matrix. However, the mechanisms that excessive mechanical forces lead to AF cell apoptosis are not clear. The present study was to look for how AF cells sense mechanical changes. In vivo experiments, the involvement of mechanoreceptors in apoptosis was examined by RT-PCR and/or immunoblotting in the lumbar spine of rats subjected to unbalanced dynamic and static forces. In vitro experiments, we investigated apoptotic signaling pathways in untransfected and transfected AF cells with the lentivirus vector for rat β1 integrin overexpression after cyclic stretch. Apoptosis in AF cells was assessed using flow cytometry, Hoechst 33258 nuclear staining. Western blotting was used to analyze expression of β1 integrin and caspase-3 and ERK1/2 MAPK signaling molecules. In the rat IVDD model, unbalanced dynamic and static forces induced apoptosis of disc cells, which corresponded to decreased expression of β1 integrin. Cyclic stretch-induced apoptosis in rat AF cells correlated with the activation of caspase-3 and with decreased levels of β1 integrin and the phosphorylation levels of ERK1/2 activation level. However, the overexpression of β1 integrin in AF cells ameliorated cyclic stretch-induced apoptosis and decreased caspase-3 activation. Furthermore, ERK1/2-specific inhibitor promotes apoptosis in vector β1-infected AF cells. These results suggest that the disruption of β1 integrin signaling may underlie disc cell apoptosis induced by mechanical stress. Further work is necessary to fully elucidate the pathophysiological mechanisms that underlie IVDD caused by unbalanced dynamic and static forces.

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Abbreviations

IVDD:

Intervertebral disc degeneration

AF:

Annulus fibrosus

BK channels:

Large-conductance Ca2+-activated K+ channels

DDR2:

Discoidin domain receptor 2

TRPV4:

Transient receptor potential vanilloid-4

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (Grant No. 81272038), Program for Innovative Research Team of Shanghai Municipal Education Commission (Phase II).

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Author notes

  1. Wei Ding

    Present address: Jiangsu Jiangyin People’ Hospital, Jiangyin, People’s Republic of China

  2. Wei Sun

    Present address: Affiliated Hospital of Xuzhou Medical College, Xuzhou, People’s Republic of China

Authors and Affiliations

  1. Shanghai Key Laboratory of Orthopaedic Implants, Department of Orthopaedics, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, 639 Zhizaoju Road, Shanghai, 200011, People’s Republic of China

    Kai Zhang, Wei Ding, Wei Sun, Xiao-jiang Sun, You-zhuan Xie, Chang-qing Zhao & Jie Zhao

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  1. Kai Zhang
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  2. Wei Ding
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  3. Wei Sun
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  6. Chang-qing Zhao
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Correspondence to Chang-qing Zhao or Jie Zhao.

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The authors declare that they have no conflict of interest.

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Dr. Kai Zhang and Wei Ding have contributed equally to this study.

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Zhang, K., Ding, W., Sun, W. et al. Beta1 integrin inhibits apoptosis induced by cyclic stretch in annulus fibrosus cells via ERK1/2 MAPK pathway. Apoptosis 21, 13–24 (2016). https://doi.org/10.1007/s10495-015-1180-7

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  • DOI: https://doi.org/10.1007/s10495-015-1180-7

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