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Calycosin Preserves BDNF/TrkB Signaling and Reduces Post-Stroke Neurological Injury after Cerebral Ischemia by Reducing Accumulation of Hypertrophic and TNF-α-Containing Microglia in Rats

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Abstract

Both brain-derived neurotrophic factor (BDNF) and microglia activation are involved in the pathogenesis of ischemic stroke. Herein, we attempt to ascertain whether Calycosin, an isoflavonoid, protects against ischemic stroke by modulating the endogenous production of BDNF and/or the microglia activation. This study was a prospective, randomized, blinded and placebo-controlled preclinical experiment. Sprague-Dawley adult rats, subjected to transient focal cerebral ischemia by middle cerebral artery occlusion (MCAO), were treated randomly with 0 (corn oil and/or saline as placebo), 30 mg/kg of Calycosin and/or 1 mg/kg of a tropomyosin-related kinase B (TrkB) receptor antagonist (ANA12) at 1 h after reperfusion and once daily for a total of 7 consecutive days. BDNF and its functional receptor, full-length TrkB (TrkB-FL) levels, the percentage of hypertrophic microglia, tumor necrosis factor-α (TNF-α)-containing microglia, and degenerative and apoptotic neurons in ischemic brain regions were determined 7 days after cerebral ischemia. A battery of functional sensorimotor test was performed over 7 days. Post-stroke Calycosin therapy increased the cerebral expression of BDNF/TrkB, ameliorated the neurological injury and switched the microglia from the activated amoeboid state to the resting ramified state in ischemic stroke rats. However, the beneficial effects of BDNF/ TrkB-mediated Calycosin could be reversed by ANA12. Our data indicate that BDNF/TrkB-mediated Calycosin ameliorates rat ischemic stroke injury by switching the microglia from the activated amoeboid state to the resting ramified state.

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Abbreviations

BDNF:

brain-derived neurotrophic factor

CBF:

cerebral blood flow

CIV:

corrected infarction volume

d :

thickness of the brain slices

DAPI:

4′,6-diamidino-2-phenylindole

ECA:

external carotid artery

ELISA:

enzyme-linked immunosorbent assay

Iba-1:

ionized calcium binding adaptor molecule 1

ICA:

internal carotid artery

LT:

left hemisphere

MCAO:

middle cerebral artery occlusion

mNSS:

modified neurological severity score

NeuN:

neuronal nuclei

PBS:

phosphate-buffered saline

RA:

Radix Astragali

RI:

right side infarct

RT:

right hemisphere

SD:

Sprague-Dawley

TNF-α:

tumor necrosis factor-alpha

TrkB-FL:

full-length tyrosine kinase receptor B

TTC:

2,3,5-triphenyl tetrazolium chloride

TUNEL:

terminal deoxynucleotidyl transferase dUTP nick end labeling

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Acknowledgments

The authors would like to thank Ms. Meng-Tsung Ho for her excellent editorial assistance in manuscript preparation.

Funding

This work was supported by grants from the Ministry of Science and Technology (Taiwan) (MOST 107-2314-B-384-004) and Chi Mei Medical Center (Tainan, Taiwan) (CMFHT10504).

Author information

C.C.H. and W.P.L. performed the MCAO surgeries and western blotting. T.W.K. performed the rat behavioral tests and immunohistochemical staining. C.C.H. and H.J.L. designed and coordinated the study. C.P.C. and H.J.L. conceived the study and wrote the manuscript. All authors read and approved the final manuscript.

Correspondence to Ching-Ping Chang or Hung-Jung Lin.

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Ethics Approval and Consent to Participate

All animal research was ethically approved by the Institutional Animal Care and Use Committee of Chi Mei Medical Center (IACUC approved no. 105110328). Experiments adhered to guidelines from the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

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Not applicable.

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The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Hsu, C., Kuo, T., Liu, W. et al. Calycosin Preserves BDNF/TrkB Signaling and Reduces Post-Stroke Neurological Injury after Cerebral Ischemia by Reducing Accumulation of Hypertrophic and TNF-α-Containing Microglia in Rats. J Neuroimmune Pharmacol (2020) doi:10.1007/s11481-019-09903-9

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Keywords

  • Stroke
  • Calycosin
  • Brain-derived neurotrophic factor
  • Microglia
  • Neuronal death