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Regulation of Autophagy and Ubiquitinated Protein Accumulation by bFGF Promotes Functional Recovery and Neural Protection in a Rat Model of Spinal Cord Injury

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Abstract

The role of autophagy in the recovery of spinal cord injury remains controversial; in particular, the mechanism of autophagy regulated degradation of ubiquitinated proteins has not been discussed to date. In this study, we investigated the protective role of basic fibroblast growth factor (bFGF) both in vivo and in vitro and demonstrated that excessive autophagy and ubiquitinated protein accumulation is involved in the rat model of trauma. bFGF administration improved recovery and increased the survival of neurons in spinal cord lesions in the rat model. The protective effect of bFGF is related to the inhibition of autophagic protein LC3II levels; bFGF treatment also enhances clearance of ubiquitinated proteins by p62, which also increases the survival of neuronal PC-12 cells. The activation of the downstream signals of the PI3K/Akt/mTOR pathway by bFGF treatment was detected both in vivo and in vitro. Combination therapy including the autophagy activator rapamycin partially abolished the protective effect of bFGF. The present study illustrates that the role of bFGF in SCI recovery is related to the inhibition of excessive autophagy and enhancement of ubiquitinated protein clearance via the activation of PI3K/Akt/mTOR signaling. Overall, our study suggests a new trend for bFGF drug development for central nervous system injuries and sheds light on protein signaling involved in bFGF action.

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Abbreviations

bFGF:

Basic fibroblast growth factor

SCI:

Spinal cord injury

LC3:

Light chain 3

mTOR:

Mammalian target rapamycin

3-MA:

3-Methyladenine

BBB:

Basso Beattie, and Bresnahan

UPS:

Ubiquitin–proteasome system

IGF-1:

Insulin-like growth factor-1

NGF:

Nerve growth factor

BDNF:

Brain-derived neurotrophic factor

GDNF:

Glial cell line-derived neurotrophic factor

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Acknowledgments

This work was supported by Zhejiang Provincial Program for the Cultivation of High-level Innovative Health talents (to J.X.), National Natural Science Funding of China (81200958, 81200010), State Key Basic Research Development Program (2012CB518105), Zhejiang Provincial Project of Key Group (2010R5004202), Ninbo Natural Science Foundation (2012A610255).

Conflict of interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. School of Pharmacy, Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical College, Wenzhou, 325035, China

    Hong-Yu Zhang, Zhou-Guang Wang, Fen-Zan Wu, Jie Yang, Bei-Bei Lin, Li Lin, Xiao-Kun Li & Jian Xiao

  2. Department of Neurology, Cixi Hospital, Wenzhou Medical College, Ningbo, 315300, China

    Zhou-Guang Wang & Chao-Shi Gan

  3. Institute of Hypoxia Medical Research, Teaching Center of Functional Experiment, Basic Medical School, Wenzhou Medical College, Wenzhou, 325035, China

    Xiao-Xia Kong

  4. Department of Orthopaedics, The Second Affiliated Hospital, Wenzhou Medical College, Wenzhou, 325000, China

    Shi-Ping Zhu & Hua-Zi Xu

  5. Institute of Basic Medical Science, Chinese PLA General Hospital, Beijing, 100853, China

    Xiao-Bing Fu

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  1. Hong-Yu Zhang
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Corresponding authors

Correspondence to Hua-Zi Xu or Jian Xiao.

Additional information

H.-Y. Zhang Z.-G. Wang and F.-Z. Wu contributed equally to this study.

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Fig. S1

(A) Statistical results of NE staining of sham, SCI model group and SCI model mice treated by bFGF group of Figure 1D. (B) Statistical results of Immunofluorecence staining results of LC3 of Figure 2A. (C) Statistical results of Immunofluorecence staining results of Ub of Figure 4C. (D) Statistical results of Immunofluorecence staining results of LC3 in PC12 cell of Figure 7B. (JPEG 69 kb)

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Zhang, HY., Wang, ZG., Wu, FZ. et al. Regulation of Autophagy and Ubiquitinated Protein Accumulation by bFGF Promotes Functional Recovery and Neural Protection in a Rat Model of Spinal Cord Injury. Mol Neurobiol 48, 452–464 (2013). https://doi.org/10.1007/s12035-013-8432-8

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