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Mn-SOD Upregulation by Electroacupuncture Attenuates Ischemic Oxidative Damage via CB1R-Mediated STAT3 Phosphorylation

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Abstract

Electroacupuncture (EA) pretreatment elicits the neuroprotective effect against cerebral ischemic injury through cannabinoid receptor type 1 receptor (CB1R). In current study, we aimed to investigate whether the signal transducer and activator of transcription 3 (STAT3) and manganese superoxide dismutase (Mn-SOD) were involved in the antioxidant effect of EA pretreatment through CB1R. At 2 h after EA pretreatment, focal cerebral ischemic injury was induced by transient middle cerebral artery occlusion for 60 min in C57BL/6 mice. The expression of Mn-SOD in the penumbra was assessed by Western blot and immunoflourescent staining at 2 h after reperfusion. In the presence or absence of Mn-SOD small interfering RNA (siRNA), the neurological deficit score, the infarct volume, the terminal deoxynucleotidyl transferase-mediated dUDP-biotin nick end labeling (TUNEL) staining, and oxidative stress were evaluated. Furthermore, the Mn-SOD protein expression and phosphorylation of STAT3 at Y705 were also determined in the presence and absence of CB1R antagonists (AM251, SR141716) and CB1R agonists (arachidonyl-2-chloroethylamide (ACEA), WIN 55,212-2). EA pretreatment upregulated the Mn-SOD protein expression and Mn-SOD-positive neuronal cells at 2 h after reperfusion. EA pretreatment also attenuated oxidative stress, inhibited cellular apoptosis, and induced neuroprotection against ischemic damage, whereas these beneficial effects of EA pretreatment were reversed by knockdown of Mn-SOD. Mn-SOD upregulation and STAT3 phosphorylation by EA pretreatment were abolished by two CB1R antagonists, while pretreatment with two CB1R agonists increased the expression of Mn-SOD and phosphorylation level of STAT3. Mn-SOD upregulation by EA attenuates ischemic oxidative damage through CB1R-mediated STAT3 phosphorylation in stroke mice, which may represent one new mechanism of EA pretreatment-induced neuroprotection against cerebral ischemia.

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Abbreviations

t-PA:

Tissue-type plasminogen activators

ROS:

Reactive oxygen species

I/R:

Ischemia/reperfusion

SOD:

Superoxide dismutase

Mn-SOD:

Manganese superoxide dismutase

Cu/Zn-SOD:

Cu/Zn superoxide dismutase

EA:

Electroacupuncture

MCAO:

Middle cerebral artery occlusion

2-AG:

2-Arachidonylglycerol

AEA:

N-Arach-idonoylethanolamine-anandamide

CB1R:

Cannabinoid receptor type 1 receptor

Trx:

Thioredoxin

GSH:

Glutathione

MDA:

Malondialdehyde

STAT3:

Signal transducer and activator of transcription 3

TUNEL:

Terminal deoxynucleotidyl transferase-mediated dUDP-biotin nick end labeling

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Acknowledgments and Funding

This work was supported by the National Natural Science Foundation of China (Grant 81072888 and 81173394), the Overseas, Hong Kong & Macao Scholars Collaborated Researching Fund (Grant 81228022), and the National Key Basic Research and Development Program (973, Grant 2014 CB543202).

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Authors and Affiliations

  1. Department of Anesthesiology Xijing Hospital, Fourth Military Medical University, Xi’an, 710032, Shaanxi Province, China

    Sisi Sun, Yang Gao, Zhaoyu Liu, Qian Zhai, Lize Xiong & Qiang Wang

  2. Department of Psychiatry, Xijing Hospital, Fourth Military Medical University, Xi’an, 710032, Shaanxi Province, China

    Min Cai

  3. Department of Physiology, Fourth Military Medical University, Xi’an, 710032, China

    Xiyao Chen

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  1. Sisi Sun
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  2. Xiyao Chen
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  3. Yang Gao
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  4. Zhaoyu Liu
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  5. Qian Zhai
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  6. Lize Xiong
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  7. Min Cai
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  8. Qiang Wang
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Correspondence to Min Cai or Qiang Wang.

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Sisi Sun and Xiyao Chen contributed to this work equally.

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Sun, S., Chen, X., Gao, Y. et al. Mn-SOD Upregulation by Electroacupuncture Attenuates Ischemic Oxidative Damage via CB1R-Mediated STAT3 Phosphorylation. Mol Neurobiol 53, 331–343 (2016). https://doi.org/10.1007/s12035-014-8971-7

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