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Necrostatin-1 Suppresses Autophagy and Apoptosis in Mice Traumatic Brain Injury Model

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Abstract

Traumatic brain injury (TBI) results in neuronal apoptosis, autophagic cell death and necroptosis. Necroptosis is a newly discovered caspases-independent programmed necrosis pathway which can be triggered by activation of death receptor. Previous works identified that necrostatin-1 (NEC-1), a specific necroptosis inhibitor, could reduce tissue damage and functional impairment through inhibiting of necroptosis process following TBI. However, the role of NEC-1 on apoptosis and autophagy after TBI is still not very clear. In this study, the amount of TBI-induced neural cell deaths were counted by PI labeling method as previously described. The expression of autophagic pathway associated proteins (Beclin-1, LC3-II, and P62) and apoptotic pathway associated proteins (Bcl-2 and caspase-3) were also respectively assessed by immunoblotting. The data showed that mice pretreated with NEC-1 reduced the amount of PI-positive cells from 12 to 48 h after TBI. Immunoblotting results showed that NEC-1 suppressed TBI-induced Beclin-1 and LC3-II activation which maintained p62 at high level. NEC-1 pretreatment also reversed TBI-induced Bcl-2 expression and caspase-3 activation, as well as the ratio of Beclin-1/Bcl-2. Both 3-MA and NEC-1 suppressed TBI-induced caspase-3 activation and LC3-II formation, Z-VAD only inhibited caspase-3 activation but increased LC3-II expression at 24 h post-TBI. All these results revealed that multiple cell death pathways participated in the development of TBI, and NEC-1 inhibited apoptosis and autophagy simultaneously. These coactions may further explain how can NEC-1 reduce TBI-induced tissue damage and functional deficits and reflect the interrelationship among necrosis, apoptosis and autophagy.

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Abbreviations

TBI:

Traumatic brain injury

NEC-1:

Necrostatin-1

i.c.v:

Intracerebroventricle

PI:

Propidium iodide

LC3:

Microtubule-associated protein- 1A/1B light chain 3

3-MA:

3-Methyladenine

Z-VAD:

Pan-caspase inhibitor Z-VAD-FMK

p62/SQSTM1:

Sequestosome 1 protein

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (30872666 and 81172911) and the Shanghai Forensic Key Lab Foundation (KF1005).

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Authors and Affiliations

  1. Department of Forensic Science and Laboratory of Neural Injury, Medical College of Soochow University, Suzhou, 215123, Jiangsu, People’s Republic of China

    Yao-Qi Wang, Long Wang, Ming-Yang Zhang, Tao Wang, Hai-Jun Bao, Wei-Li Liu, Ding-Kun Dai, Lu Zhang, Pan Chang, Wen-Wen Dong, Xi-Ping Chen & Lu-Yang Tao

  2. Department of Forensic Science, Medical College of Nantong University, Nantong, 226001, Jiangsu, People’s Republic of China

    Ming-Yang Zhang

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  1. Yao-Qi Wang
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  2. Long Wang
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Correspondence to Xi-Ping Chen or Lu-Yang Tao.

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Yao-Qi Wang and Long Wang contributed equally to this work.

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Wang, YQ., Wang, L., Zhang, MY. et al. Necrostatin-1 Suppresses Autophagy and Apoptosis in Mice Traumatic Brain Injury Model. Neurochem Res 37, 1849–1858 (2012). https://doi.org/10.1007/s11064-012-0791-4

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