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Activation of Melanocortin 1 Receptor Attenuates Early Brain Injury in a Rat Model of Subarachnoid Hemorrhage viathe Suppression of Neuroinflammation through AMPK/TBK1/NF-κB Pathway in Rats

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Abstract

Neuroinflammation plays a vital role in early brain injury (EBI) following subarachnoid hemorrhage (SAH). The hypothesis of this study was that activation of melanocortin 1 receptor (MC1R) with BMS-470539 attenuates EBI by suppression of neuroinflammation after SAH. We utilized BMS-470539, MSG-606, and MRT-68601 to verify the neuroprotective effects of MC1R. We evaluated brain water content, short-term and long-term neurobehavior after SAH. Western blotting and immunofluorescence staining were utilized to assess the changes of protein levels. The results of western blotting suggested that the expressions of MC1R, phosphorylated-adenosine monophosphate-activated protein kinase (p-AMPK), and phosphorylated-TANK binding kinase 1 (p-TBK1) were increased and reached their peak points at 24 h following SAH. Moreover, BMS-470539 treatment notably attenuated neurological deficits caused by SAH, and also notably improved long-term spatial learning and memory abilities after SAH. The underlying mechanisms of the neuroprotection of BMS-470539 involved the suppression of microglia activation, promotion of CD206+ microglia transformation and reduction of neutrophil infiltration by increasing the levels of p-AMPK and p-TBK1 while decreasing the levels of NF-κB, IL-1β, and TNFα. The neuroprotective effects of BMS-470539 were significantly abolished by MSG-606 and MRT-68601. The activation of MC1R with BMS-470539 notably attenuates EBI after SAH by suppression of microglial activation and neutrophil infiltration via the AMPK/TBK1/NF-κB signaling pathway.

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Abbreviations

EBI :

Early brain injury

SAH :

Subarachnoid hemorrhage

MC1R :

Melanocortin 1 receptor

AMPK :

Adenosine monophosphate-activated protein kinase

TBK1 :

TANK-binding kinase 1

CNS :

Central nervous system

IKK :

IκB kinase β

SD :

Sprague–Dawley

MPO :

Myeloperoxidase

GFAP :

Glial fibrillary acidic protein

LKB :

Liver kinase B

CaMKK-β :

Ca2+/calmodulin-dependent protein kinase kinase-β

TAK1 :

Transforming growth factor-β-activated kinase 1

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Acknowledgements

This work was supported by the National Institutes of Health (Grants NS081740 and NS082184); National Key Research and Development Program of China (2017YFC1308500), Key Program of Science and Technology Development of Zhejiang (2017C03021); Grant WKJ-ZJ-1615 (2016149634) from a major science and technology project in medical and health of Zhejiang province. Basic Public Interest Research Plan of Zhejiang Province (LY18H090001).

Author Contributions Statement

This study was designed by WX, JM, JT, JZ, and JHZ. The experiments were completed by WX, JM, UO, ZDT, TZ, PW, TL, and YZ. JP, BE and AS performed statistical analysis. WX and JM finished writing the manuscript. JZ and JHZ provided the supervision and final check. All the authors read the final version of this paper and approved it.

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Correspondence to Jianmin Zhang or John H. Zhang.

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Ethical Approval and Consent to Participate

All animal experiments were performed according to the Institutional Animal Care and Use Committee (IACUC) at Loma Linda University. The procedures were conducted according to the National Institutes of Health’s Guide for the Care and the Use of Laboratory Animals and the ARRIVE (Animal Research: Reporting In Vivo Experiments) guidelines.

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The datasets analyzed during the present study are available from the corresponding author on reasonable request.

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The authors report no conflicts of interest.

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Xu, W., Mo, J., Ocak, U. et al. Activation of Melanocortin 1 Receptor Attenuates Early Brain Injury in a Rat Model of Subarachnoid Hemorrhage viathe Suppression of Neuroinflammation through AMPK/TBK1/NF-κB Pathway in Rats. Neurotherapeutics 17, 294–308 (2020). https://doi.org/10.1007/s13311-019-00772-x

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Keywords

  • Early brain injury
  • Subarachnoid hemorrhage
  • Melanocortin 1 Receptor
  • TBK1
  • Neuroinflammation