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Anti-Necroptosis Chemical Necrostatin-1 Can Also Suppress Apoptotic and Autophagic Pathway to Exert Neuroprotective Effect in Mice Intracerebral Hemorrhage Model

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Abstract

Necroptosis was recently discovered as one form of programmed cell death (PCD) and could be specifically inhibited by necrostatin-1. The aim of this study was to examine the effect of necrostatin-1 on brain injury and investigate the role of necrostatin-1 on the other two types PCD (apoptosis and autophagic cell death) in a mouse intracerebral hemorrhage (ICH) model. Male ICR mice received an infusion of type IV collagenase to induce ICH or saline as control into the left striatum. In the presence of vehicle, 3-MA, zVAD, and necrostatin-1 were pretreated with a single intracerebroventricular (i.c.v.) injection in the ipsilateral ventricle 15 min before ICH, respectively. Compared with vehicle groups, necrostatin-1 treatment significantly reduced injury volume and propidium iodide-positive cells at 24 and 72 h after ICH. Immunoblotting analysis showed that necrostatin-1 treatment suppressed autophagic-associated proteins (LC3-II, Beclin-1) and maintained p62 at normal level at 24 and 72 h after ICH. In addition, necrostatin-1 treatment enhanced the protein level of Bcl-2 and decreased the protein level of cleaved caspase-3 and the Beclin-1/Bcl-2 ratio at 24 and 72 h after ICH. Moreover, both 3-MA and necrostatin-1 treatment could suppress cleaved caspase-3 and LC3-II production, whereas zVAD treatment could inhibit caspase-3 cleavage but increased LC3-II protein levels at 72 h after ICH. Taken together, the data demonstrated for the first time that the specific inhibitor necrostatin-1 suppressed apoptosis and autophagy to exert these neuroprotective effects after ICH and that there existed a cross-talk among necroptosis, apoptosis, and autophagy after ICH.

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Abbreviations

PCD:

Programmed cell death

ICH:

Intracerebral hemorrhage

3-MA:

3-Methyladenine

zVAD:

Pan-caspase inhibitor Z-VAD-FMK

i.c.v.:

Intracerebroventricular

PI:

Propidium iodide

LC3:

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

p62/SQSTM1:

Sequestosome 1 protein

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (no. 81271379, 81373251, no. 81172911, no. 81301039), and the science and technology development project of Suzhou (no. SZP201304).

Conflict of Interest

There is no conflict of interest to disclose.

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Authors and Affiliations

  1. Institute of Forensic Sciences, Soochow University, Suzhou, 215123, Jiangsu, China

    Pan Chang, Wenwen Dong, Mingyang Zhang, Zufeng Wang, Yaoqi Wang, Tao Wang, Yuan Gao, Huanhuan Meng, Bin Luo, Chengliang Luo, Xiping Chen & Luyang Tao

  2. Department of Forensic Sciences, Medical College of Nantong University, Nantong, 226001, Jiangsu, China

    Mingyang Zhang

Authors
  1. Pan Chang
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  2. Wenwen Dong
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  3. Mingyang Zhang
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  4. Zufeng Wang
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  5. Yaoqi Wang
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  6. Tao Wang
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  7. Yuan Gao
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  9. Bin Luo
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  11. Xiping Chen
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Correspondence to Xiping Chen or Luyang Tao.

Additional information

Pan Chang, Wenwen Dong, and Mingyang Zhang contributed equally to this work.

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Chang, P., Dong, W., Zhang, M. et al. Anti-Necroptosis Chemical Necrostatin-1 Can Also Suppress Apoptotic and Autophagic Pathway to Exert Neuroprotective Effect in Mice Intracerebral Hemorrhage Model. J Mol Neurosci 52, 242–249 (2014). https://doi.org/10.1007/s12031-013-0132-3

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  • DOI: https://doi.org/10.1007/s12031-013-0132-3

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