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).
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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.
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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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Hsu, CC., Kuo, TW., Liu, WP. 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 15, 326–339 (2020). https://doi.org/10.1007/s11481-019-09903-9
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DOI: https://doi.org/10.1007/s11481-019-09903-9