Astragaloside VI Promotes Neural Stem Cell Proliferation and Enhances Neurological Function Recovery in Transient Cerebral Ischemic Injury via Activating EGFR/MAPK Signaling Cascades
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Radix Astragali (AR) is a commonly used medicinal herb for post-stroke disability in Traditional Chinese Medicine but its active compounds for promoting neurogenic effects are largely unknown. In the present study, we tested the hypothesis that Astragaloside VI could be a promising active compound from AR for adult neurogenesis and brain repair via targeting epidermal growth factor (EGF)-mediated MAPK signaling pathway in post-stroke treatment. By using cultured neural stem cells (NSCs) and experimental stroke rat model, we investigated the effects of Astragaloside VI on inducing NSCs proliferation and self-renewal in vitro, and enhancing neurogenesis for the recovery of the neurological functions in post-ischemic brains in vivo. For animal experiments, rats were undergone 1.5 h middle cerebral artery occlusion (MCAO) plus 7 days reperfusion. Astragaloside VI (2 μg/kg) was daily administrated by intravenous injection (i.v.) for 7 days. Astragaloside VI treatment promoted neurogenesis and astrogenic formation in dentate gyrus zone, subventricular zone, and cortex of the transient ischemic rat brains in vivo. Astragaloside VI treatment enhanced NSCs self-renewal and proliferation in the cultured NSCs in vitro without affecting NSCs differentiation. Western blot analysis showed that Astragaloside VI up-regulated the expression of nestin, p-EGFR and p-MAPK, and increased neurosphere sizes, whose effects were abolished by the co-treatment of EGF receptor inhibitor gefitinib and ERK inhibitor PD98059. Behavior tests revealed that Astragaloside VI promoted the spatial learning and memory and improved the impaired motor function in transient cerebral ischemic rats. Taken together, Astragaloside VI could effectively activate EGFR/MAPK signaling cascades, promote NSCs proliferation and neurogenesis in transient cerebral ischemic brains, and improve the repair of neurological functions in post-ischemic stroke rats. Astragaloside VI could be a new therapeutic drug candidate for post-stroke treatment.
KeywordsAstragaloside VI Transient cerebral ischemia Neural stem cell Proliferation Epidermal growth factor
Epidermal growth factor
Neural stem cells
Middle cerebral artery occlusion
Traditional Chinese medicine
We appreciate Professor Guowei Qin from Shanghai Institute of Meteria Medica for providing the sample of Astrogaloside VI for our study.
The work was performed and accomplished by all authors. S-JG and XC received fund and designed the experiments. XC, HW, and C-HS conducted the experiments and statistical analyses. X-XJ and QW provided technical support for animal experiments, experimental design, data interpretation, and gave comments on the manuscript. XC and S-JG wrote the manuscript. All authors have read and approved the final manuscript.
This study was supported by the grants from Shenzhen Science and Technology Innovation Commission (JCYJ20150402152005623), 2011 State Key Project of National Natural Foundation of China (No. 81630101,SIRI/04/09/2014/2), and National Natural Foundation of China (No. 81703741).
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
Animal experimental protocols were conducted in accordance with the national and institutional guidelines on ethics and biosafety, which were approved and regulated by the Committee on the Use of Live Animals in Teaching and Research (CULATR), HKU. This article does not contain any studies with human participants performed by any of the authors.
- 3.Seo TB, Kim TW, Shin MS, Ji ES, Cho HS, Lee JM, Kim TW, Kim CJ (2014) Aerobic exercise alleviates ischemia-induced memory impairment by enhancing cell proliferation and suppressing neuronal apoptosis in hippocampus. Int Neurourol J 18(4):187–197. https://doi.org/10.5213/inj.2014.18.4.187 CrossRefPubMedPubMedCentralGoogle Scholar
- 6.Tian L, Nie H, Zhang Y, Chen Y, Peng Z, Cai M et al (2014) Recombinant human thioredoxin-1 promotes neurogenesis and facilitates cognitive recovery following cerebral ischemia in mice. Neuropharmacology 77:453–464. https://doi.org/10.1016/j.neuropharm.2013.10.027.1 CrossRefPubMedGoogle Scholar
- 7.Cittolin-Santos GF, de Assis AM, Guazzelli PA, Paniz LG, da Silva JS, Calcagnotto ME, Hansel G, Zenki KC et al (2016) Guanosine exerts neuroprotective effect in an experimental model of acute ammonia intoxication. Mol Neurobiol 54:3137–3148. https://doi.org/10.1007/s12035-016-9892-4 CrossRefPubMedGoogle Scholar
- 21.Morita M, Kozuka N, Itofusa R, Yukawa M, Kudo Y (2005) Autocrine activation of EGF receptor promotes oscillation of glutamate-induced calcium increase in astrocytes cultured in rat cerebral cortex. J Neurochem 95(3):871–879. https://doi.org/10.1111/j.1471-4159.2005.03430.x CrossRefPubMedGoogle Scholar
- 22.Jori FP, Galderisi U, Piegari E, Cipollaro M, Cascino A, Peluso G, Cotrufo R, Giordano A et al (2003) EGF-responsive rat neural stem cells: molecular follow-up of neuron and astrocyte differentiation in vitro. J Cell Physiol 195(2):220–233. https://doi.org/10.1002/jcp.10249 CrossRefPubMedGoogle Scholar
- 23.Anderson NG, Ahmad T, Chan K, Dobson R, Bundred NJ (2001) ZD1839 (Iressa), a novel epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, potently inhibits the growth of EGFR-positive cancer cell lines with or without erbB2 overexpression. Int J Cancer 94(6):774–782CrossRefPubMedGoogle Scholar
- 32.Sutterlin P, Williams EJ, Chambers D, Saraf K, von Schack D, Reisenberg M, Doherty P, Williams G (2013) The molecular basis of the cooperation between EGF, FGF and eCB receptors in the regulation of neural stem cell function. Mol Cell Neurosci 52:20–30. https://doi.org/10.1016/j.mcn.2012.10.006 CrossRefPubMedGoogle Scholar
- 38.Hao CZ, Wu F, Shen J, Lu L, Fu DL, Liao WJ, Zheng GQ (2012) Clinical efficacy and safety of buyang huanwu decoction for acute ischemic stroke: a systematic review and meta-analysis of 19 randomized controlled trials. Evid Based Complement Alternat Med 2012:630124–630110. https://doi.org/10.1155/2012/630124 CrossRefPubMedPubMedCentralGoogle Scholar
- 41.Fu S, Gu Y, Jiang J-Q, Chen X, Mingjing X, Chen X, Shen J (2014) Calycosin-7-O-beta-D-glucoside regulates nitric oxide /caveolin-1/matrix metalloproteinases pathway and protects blood-brain barrier integrity in experimental cerebral ischemia-reperfusion injury. J Ethnopharmacol 155(1):692–701CrossRefPubMedGoogle Scholar
- 43.Qu YZ, Li M, Zhao YL, Zhao ZW, Wei XY, Liu JP, Gao L, Gao GD (2009) Astragaloside IV attenuates cerebral ischemia-reperfusion-induced increase in permeability of the blood-brain barrier in rats. Eur J Pharmacol 606(1–3):137–141. https://doi.org/10.1016/j.ejphar.2009.01.022 CrossRefPubMedGoogle Scholar
- 45.Li M, Qu YZ, Zhao ZW, Wu SX, Liu YY, Wei XY, Gao L, Gao GD (2012) Astragaloside IV protects against focal cerebral ischemia/reperfusion injury correlating to suppression of neutrophils adhesion-related molecules. Neurochem Int 60(5):458–465. https://doi.org/10.1016/j.neuint.2012.01.026 CrossRefPubMedGoogle Scholar
- 49.Squire LR, Zola SM (1996) Ischemic brain damage and memory impairment: a commentary. Hippocampus 6(5):546–552. https://doi.org/10.1002/(SICI)1098-1063(1996)6:5<546::AID-HIPO7>3.0.CO;2-G CrossRefPubMedGoogle Scholar
- 55.Justicia C, Planas AM (1999) Transforming growth factor-alpha acting at the epidermal growth factor receptor reduces infarct volume after permanent middle cerebral artery occlusion in rats. J Cereb Blood Flow Metab 19(2):128–132. https://doi.org/10.1097/00004647-199902000-00002 CrossRefPubMedGoogle Scholar
- 59.Yang K, Cao F, Sheikh AM, Malik M, Wen G, Wei H, Ted Brown W, Li X (2013) Up-regulation of Ras/Raf/ERK1/2 signaling impairs cultured neuronal cell migration, neurogenesis, synapse formation, and dendritic spine development. Brain Struct Funct 218(3):669–682. https://doi.org/10.1007/s00429-012-0420-7 CrossRefPubMedGoogle Scholar
- 61.Rojewska E, Popiolek-Barczyk K, Kolosowska N, Piotrowska A, Zychowska M, Makuch W, Przewlocka B, Mika J (2015) PD98059 influences immune factors and enhances opioid analgesia in model of neuropathy. PLoS One 10(10):e0138583. https://doi.org/10.1371/journal.pone.0138583 eCollection 2015CrossRefPubMedPubMedCentralGoogle Scholar