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Non-invasive Vagus Nerve Stimulation Protects Against Cerebral Ischemia/Reperfusion Injury and Promotes Microglial M2 Polarization Via Interleukin-17A Inhibition

  • Xiao-Ping Zhao
  • Yuan Zhao
  • Xiao-Ya Qin
  • Li-Yuan Wan
  • Xiao-Xuan FanEmail author
Article
  • 79 Downloads

Abstract

Microglia play an essential role during cerebral an ischemia/reperfusion (I/R)-related inflammatory process. Because the M2 phenotype of microglia exhibits anti-inflammation activity, it has become a promising target for anti-inflammatory therapy. Vagus nerve stimulation (VNS) reportedly has neuroprotective effects against cerebral I/R injuries via its anti-inflammatory action. The aim of this study was to investigate the ability of non-invasive VNS (nVNS) to alleviate cerebral I/R in mice by promoting microglial M2 polarization. Neurological scoring and cerebral infarct volume assessments were performed 72 h after a middle cerebral artery occlusion (MCAO)-induced stroke. M2 phenotype microglia were identified by immunohistochemistry staining using Arg-1 and Iba-1 antibodies. The protein expressions of Arg-1, IL-17A, IL-10, Bax, and Bcl-2 were detected by Western blot. Apoptotic cells were detected using TUNEL staining. According to our results, nVNS decreased infarct volume, improved neurological outcomes, reduced apoptotic neurons (TUNEL+NeuN+ cells), and promoted microglial M2 polarization as indicated by elevated Arg-1 protein expression and increased Arg-1+ cells after MCAO. Moreover, nVNS attenuated the increased levels of IL-17A protein expression after MCAO. To test the possible involvement of IL-17A in nVNS-induced neuroprotection and microglial M2 polarization, 1-μg recombinant IL-17A (rIL-17A) was intranasally administered once daily for three consecutive days after reperfusion. We found that the intranasal administration of rIL-17A nullified the nVNS-induced promotion of microglial M2 polarization. Furthermore, rIL-17A administration abolished the neuroprotective effect of nVNS. In conclusion, our study identifies microglial M2 polarization as an important mechanism underlying the nVNS-mediated neuroprotection against cerebral I/R. This effect of nVNS could be attributed to the inhibition of IL-17A expression.

Keywords

Cerebral ischemia Microglia Vagus nerve stimulation IL-17A MCAO 

Abbreviations

I/R

Ischemia/reperfusion

Arg-1

Arginase 1

CCA

The common carotid artery

ICA

The middle cerebral artery

IL-17A

Interleukin-17A

MCAO

Middle cerebral artery occlusion

nVNS

Non-invasive VNS

rCBF

The regional cerebral blood flow

rIL-17A

The recombinant interleukin-17A

TTC

2,3,5-Triphenyltetrazolium chloride

TUNEL

Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling

VNS

Vagus nerve stimulation

α7AchR

α7 Acetylcholine receptor

Notes

Conflict of Interest

The authors declare that they have no conflict of interest.

Compliance with Ethical Standards

The experimental protocol was approved by the Ethics Committee and Animal Experimentation and was performed according to the Guidelines for Animal Experimentation of the Fourth Military Medical University. The experiments were carried out in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals (NIH Publications no. 80-23) revised 1996. Efforts were made to minimize animal sufferings and the number of animals used in the study.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of NeurosurgeryAffiliated Hospital of Shaanxi University of Chinese MedicineXianyangChina
  2. 2.College of foreign languagesShaanxi University of Chinese MedicineXianyangChina
  3. 3.First Clinical Medical College of Shaanxi University of Chinese MedicineXianyangChina

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