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Silibinin Prevents Autophagic Cell Death upon Oxidative Stress in Cortical Neurons and Cerebral Ischemia-Reperfusion Injury

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Abstract

Neuronal apoptosis and oxidative stress are involved in most of the neurodegenerative diseases, promoting neuron survival is critical for therapy. Silibinin (SLB), which is derived from the seeds of Silybinisus laborinum L., has been widely used as an antioxidant. Here we tested the neuroprotective effects of SLB and the involved molecular mechanisms. We demonstrated that SLB promoted neuron viability upon hydrogen peroxide (H2O2) challenge and reduced hypoxia/ischemia injury in the middle cerebral artery occlusion (MCAO) mouse model. SLB reversed the decreased level of procaspase-3 and balanced Bcl-2 and Bax expression upon H2O2 insult to inhibit cell apoptosis. Furthermore, SLB suppressed the activation of autophagy by decreasing microtubule-associated protein 1 light chain 3 (LC3-II) and Beclin-1 levels under oxidative stress accordingly. SLB phosphorylated protein kinase B (Akt-1) at Ser473 in a time- and dose-dependent manner. The inhibitor for phosphoinositide-3-kinase (PI3K) wortmannin abrogated SLB-induced phosphorylation of Akt-1 and mTOR, decreased the suppression of autophagy, and therefore abolished SLB-mediated neuroprotection. All the data suggested that SLB protected neurons by inhibiting both the mitochondrial and autophagic cell death pathways. This study opens new avenues for the use of SLB in treatment of central nervous system (CNS) diseases in which oxidative stress plays a major role in disease pathogenesis. Given that it occurs naturally with low toxicity and pleiotropic effects that benefit the nervous system, SLB acts potentially as a novel therapy for ischemic injury.

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Abbreviations

CNS:

Central nervous system

DMEM/F12:

Dulbecco’s modified Eagle’s medium/F12

DMSO:

Dimethyl sulfoxide

HBSS:

Hank’s balanced salt solution

H2O2 :

Hydrogen peroxide

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

PI:

Propidium iodide

Bcl-2:

B cell lymphoma 2 protein

Bax:

Bcl-2-associated X protein

PI3K/Akt-1:

Phosphoinositide-3-kinase/protein kinase B

LC3:

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

mTOR:

Mammalian target of rapamycin

MCAO:

Middle cerebral artery occlusion

TTC:

2,3,5-Triphenyltetrazolium chloride

PVDF:

Polyvinylidene difluoride

ANOVA:

Analysis of variance

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Acknowledgments

This work was supported financially by National Natural Science Foundation of China No. 81101457 and 81371322 (to Dr. Wu) and Scientific Research Foundation for the Returned Overseas Chinese Scholars No. HG3402 (to Dr. Wu).

Conflict of interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Department of Pharmacology, School of Pharmacy, The Fourth Military Medical University, Xi’an, Shaanxi Province, 710032, People’s Republic of China

    Min Wang, Yu-Jiao Li, Hui-Nan Zhang, Ting Sun, Kun Zhang, Le Yang, Yan-Yan Guo, Shui-Bing Liu, Ming-Gao Zhao & Yu-Mei Wu

  2. Department of Pharmacy, Xijing Hospital, the Fourth Military Medical University, Xi’an, Shaanxi Province, 710032, People’s Republic of China

    Yi Ding

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  1. Min Wang
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  2. Yu-Jiao Li
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  7. Le Yang
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Corresponding authors

Correspondence to Ming-Gao Zhao or Yu-Mei Wu.

Additional information

Min Wang and Yu-Jiao Li contributed equally to this study.

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Wang, M., Li, YJ., Ding, Y. et al. Silibinin Prevents Autophagic Cell Death upon Oxidative Stress in Cortical Neurons and Cerebral Ischemia-Reperfusion Injury. Mol Neurobiol 53, 932–943 (2016). https://doi.org/10.1007/s12035-014-9062-5

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