Abstract
Genistein (Gen), as a functional food in human diet, has shown many beneficial effects on neurodegenerative diseases such as Alzheimer’s disease (AD). But the neuroprotective mechanism of Gen is not clear. Because synaptic failure is considered as the earliest phase in the pathogenesis of AD, we try to validate our hypothesis that synapse may be one target of Gen on protecting neurons. In this study, SH-SY5Y cells were pre-incubated with or without Gen for 2 h followed by the incubation with Aβ25–35 (25 μmol/L) for another 24 h. Flow cytometry, Western Blots, and RT-PCR analysis were used to test the synaptic factors. The data showed that Gen pre-treatment could reverse the Aβ25–35-induced down-regulation of synaptophysin and postsynaptic marker postsynaptic density-95. In addition, the down-regulation of NR1 and NR2B induced by Aβ25-35 which are subunits of N-methyl-d-aspartate receptor also could be antagonized by pre-treatment of Gen. Moreover, the factors of CaMKII/CREB signaling pathway were detected. The results showed that mRNA and protein expressions of (Ca2+)/calmodulin(CaM), CaMKII/pCaMKII, and CREB/pCREB were significantly down-regulated by Aβ25–35, but they were all restored by the pre-treatment of Gen. Furthermore, Gen also maintained the intracellular Ca2+ concentration which was disturbed by Aβ25–35. In conclusion, these results suggested that Gen could protect synaptic dysfunction induced by Aβ, and the mechanism might be associated with the regulation of synaptic markers and Ca2+ level through activating CaM/CaMK/CREB signaling pathway.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- Aβ:
-
β-Amyloid peptides
- AD:
-
Alzheimer’s disease
- SYN:
-
Synaptophysin
- PSD-95:
-
Postsynaptic density-95
- NMDAR:
-
N-Methyl-d-aspartate receptor
- CaM:
-
Calmodulin
- CaMKII:
-
Ca2+/calmodulin-dependent protein kinase II
- CREB:
-
cAMP response element binding protein
References
Ahmed T, Enam SA, Gilani AH (2010) Curcuminoids enhance memory in an amyloid-infused rat model of Alzheimer’s disease. Neuroscience 169(3):1296–1306. doi:10.1016/j.neuroscience.2010.05.078
Alvarez-Florez F, Vidal D, Simon E (2013) MAP-kinase activity in etiolated Cucumis sativus cotyledons: the effect of red and far-red light irradiation. Plant Physiol Biochem 63:1–7. doi:10.1016/j.plaphy.2012.11.008
Bagheri M, Roghani M, Joghataei MT, Mohseni S (2012) Genistein inhibits aggregation of exogenous amyloid-beta(1)(-)(4)(0) and alleviates astrogliosis in the hippocampus of rats. Brain Res 1429:145–154. doi:10.1016/j.brainres.2011.10.020
Bossuyt J, Bers DM (2013) Visualizing CaMKII and CaM activity: a paradigm of compartmentalized signaling. J Mol Med (Berl) 91(8):907–916. doi:10.1007/s00109-013-1060-y
Chen X, Winters C, Azzam R, Li X, Galbraith JA, Leapman RD, Reese TS (2008) Organization of the core structure of the postsynaptic density. Proc Natl Acad Sci USA 105(11):4453–4458. doi:10.1073/pnas.0800897105
Ding B, Yuan L, Yu H, Li L, Ma W, Bi Y, Feng J, Xiao R (2011) Genistein and folic acid prevent oxidative injury induced by beta-amyloid peptide. Basic Clin Pharmacol Toxicol 108(5):333–340. doi:10.1111/j.1742-7843.2010.00661.x
Ding J, Xi YD, Zhang DD, Zhao X, Liu JM, Li CQ, Han J, Xiao R (2013) Soybean isoflavone ameliorates beta-amyloid 1-42-induced learning and memory deficit in rats by protecting synaptic structure and function. Synapse 67(12):856–864. doi:10.1002/syn.21692
Feng JF, He LL, Li D, Yuan LH, Yu HL, Ma WW, Yang Y, Xi YD, Ding J, Xiao YX, Xiao R (2012) Antagonizing effects of soybean isoflavones on beta-amyloid peptide-induced oxidative damage in neuron mitochondria of rats. Basic Clin Pharmacol Toxicol 111(4):248–253. doi:10.1111/j.1742-7843.2012.00900.x
Glantz LA, Gilmore JH, Hamer RM, Lieberman JA, Jarskog LF (2007) Synaptophysin and postsynaptic density protein 95 in the human prefrontal cortex from mid-gestation into early adulthood. Neuroscience 149(3):582–591. doi:10.1016/j.neuroscience.2007.06.036
Goedert M, Spillantini MG (2006) A century of Alzheimer’s disease. Science 314(5800):777–781. doi:10.1126/science.1132814
Gong B, Pan Y, Zhao W, Knable L, Vempati P, Begum S, Ho L, Wang J, Yemul S, Barnum S, Bilski A, Gong BY, Pasinetti GM (2013) IVIG immunotherapy protects against synaptic dysfunction in Alzheimer’s disease through complement anaphylatoxin C5a-mediated AMPA-CREB-C/EBP signaling pathway. Mol Immunol 56(4):619–629. doi:10.1016/j.molimm.2013.06.016
Kim YH, Kim YS, Kang SS, Cho GJ, Choi WS (2010) Resveratrol inhibits neuronal apoptosis and elevated Ca2+/calmodulin-dependent protein kinase II activity in diabetic mouse retina. Diabetes 59(7):1825–1835. doi:10.2337/db09-1431
Lardner AL (2014) Neurobiological effects of the green tea constituent theanine and its potential role in the treatment of psychiatric and neurodegenerative disorders. Nutr Neurosci 17(4):145–155. doi:10.1179/1476830513Y.0000000079
Lazzari C, Kipanyula MJ, Agostini M, Pozzan T, Fasolato C (2015) Abeta42 oligomers selectively disrupt neuronal calcium release. Neurobiol Aging 36(2):877–885. doi:10.1016/j.neurobiolaging.2014.10.020
Li S, Jin M, Koeglsperger T, Shepardson NE, Shankar GM, Selkoe DJ (2011) Soluble Abeta oligomers inhibit long-term potentiation through a mechanism involving excessive activation of extrasynaptic NR2B-containing NMDA receptors. J Neurosci 31(18):6627–6638. doi:10.1523/JNEUROSCI.0203-11.2011
Liang R, Liu X, Wei L, Wang W, Zheng P, Yan X, Zhao Y, Liu L, Cao X (2012) The modulation of the excitability of primary sensory neurons by Ca(2)(+)-CaM-CaMKII pathway. Neurol Sci 33(5):1083–1093. doi:10.1007/s10072-011-0907-7
Liao W, Jin G, Zhao M, Yang H (2013) The effect of genistein on the content and activity of alpha- and beta-secretase and protein kinase C in Abeta-injured hippocampal neurons. Basic Clin Pharmacol Toxicol 112(3):182–185. doi:10.1111/bcpt.12009
Lin CW, Chou YE, Chiou HL, Chen MK, Yang WE, Hsieh MJ, Yang SF (2014) Pterostilbene suppresses oral cancer cell invasion by inhibiting MMP-2 expression. Expert Opin Ther Target 18(10):1109–1120. doi:10.1517/14728222.2014.947962
Liu J, Chang L, Roselli F, Almeida OF, Gao X, Wang X, Yew DT, Wu Y (2010) Amyloid-beta induces caspase-dependent loss of PSD-95 and synaptophysin through NMDA receptors. J Alzheimer’s Dis 22(2):541–556. doi:10.3233/JAD-2010-100948
Luo S, Lan T, Liao W, Zhao M, Yang H (2012) Genistein inhibits Abeta(2)(5)(-)(3)(5)-induced neurotoxicity in PC12 cells via PKC signaling pathway. Neurochem Res 37(12):2787–2794. doi:10.1007/s11064-012-0872-4
Ma WW, Hou CC, Zhou X, Yu HL, Xi YD, Ding J, Zhao X, Xiao R (2013) Genistein alleviates the mitochondria-targeted DNA damage induced by beta-amyloid peptides 25–35 in C6 glioma cells. Neurochem Res 38(7):1315–1323. doi:10.1007/s11064-013-1019-y
Nonaka M, Fujii H, Kim R, Kawashima T, Okuno H, Bito H (2014) Untangling the two-way signalling route from synapses to the nucleus, and from the nucleus back to the synapses. Philos Trans R Soc Lond B Biol Sci 369(1633):20130150. doi:10.1098/rstb.2013.0150
Pan M, Li Z, Yeung V, Xu RJ (2010) Dietary supplementation of soy germ phytoestrogens or estradiol improves spatial memory performance and increases gene expression of BDNF, TrkB receptor and synaptic factors in ovariectomized rats. Nutr Metab 7:75. doi:10.1186/1743-7075-7-75
Paula-Lima AC, Brito-Moreira J, Ferreira ST (2013) Deregulation of excitatory neurotransmission underlying synapse failure in Alzheimer’s disease. J Neurochem 126(2):191–202. doi:10.1111/jnc.12304
Querfurth HW, LaFerla FM (2010) Alzheimer’s disease. N Engl J Med 362(4):329–344. doi:10.1056/NEJMra0909142
Ripoli C, Piacentini R, Riccardi E, Leone L, Li Puma DD, Bitan G, Grassi C (2013) Effects of different amyloid beta-protein analogues on synaptic function. Neurobiol Aging 34(4):1032–1044. doi:10.1016/j.neurobiolaging.2012.06.027
Ryan TJ, Grant SG (2009) The origin and evolution of synapses. Nat Rev Neurosci 10(10):701–712. doi:10.1038/nrn2717
Selkoe DJ (2002) Alzheimer’s disease is a synaptic failure. Science 298(5594):789–791. doi:10.1126/science.1074069
Shao CY, Mirra SS, Sait HB, Sacktor TC, Sigurdsson EM (2011) Postsynaptic degeneration as revealed by PSD-95 reduction occurs after advanced Abeta and tau pathology in transgenic mouse models of Alzheimer’s disease. Acta Neuropathol 122(3):285–292. doi:10.1007/s00401-011-0843-x
Szasz BK, Lenkey N, Barth AM, Mike A, Somogyvari Z, Farkas O, Lendvai B (2008) Converging effects of Ginkgo biloba extract at the level of transmitter release, NMDA and sodium currents and dendritic spikes. Planta Med 74(10):1235–1239. doi:10.1055/s-2008-1081292
Xi YD, Yu HL, Ma WW, Ding BJ, Ding J, Yuan LH, Feng JF, Xiao R (2011) Genistein inhibits mitochondrial-targeted oxidative damage induced by beta-amyloid peptide 25-35 in PC12 cells. J Bioenergy Biomembr 43(4):399–407. doi:10.1007/s10863-011-9362-7
Xi YD, Yu HL, Ding J, Ma WW, Yuan LH, Feng JF, Xiao YX, Xiao R (2012) Flavonoids protect cerebrovascular endothelial cells through Nrf2 and PI3K from beta-amyloid peptide-induced oxidative damage. Curr Neurovasc Res 9(1):32–41
Yang J, Wu X, Yu H, Liao X, Teng L (2014) NMDA receptor-mediated neuroprotective effect of the Scutellaria baicalensis Georgi extract on the excitotoxic neuronal cell death in primary rat cortical cell cultures. Sci World J 2014:459549. doi:10.1155/2014/459549
Yu HL, Li L, Zhang XH, Xiang L, Zhang J, Feng JF, Xiao R (2009) Neuroprotective effects of genistein and folic acid on apoptosis of rat cultured cortical neurons induced by beta-amyloid 31-35. Br J Nutr 102(5):655–662. doi:10.1017/S0007114509243042
Zheng CY, Seabold GK, Horak M, Petralia RS (2011) MAGUKs, synaptic development, and synaptic plasticity. Neuroscientist 17(5):493–512. doi:10.1177/1073858410386384
Acknowledgments
This work was supported by National Natural Science Foundation of China (Nos. 81172661, 81302427 and 81330065), National High Technology Research and Development Program (863 Program) of China (No. 2010AA023003).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that there are no conflicts of interest.
Additional information
Yuan-Di Xi and Dan-Di Zhang have contributed equally to this work.
Rights and permissions
About this article
Cite this article
Xi, YD., Zhang, DD., Ding, J. et al. Genistein Inhibits Aβ25–35-Induced Synaptic Toxicity and Regulates CaMKII/CREB Pathway in SH-SY5Y Cells. Cell Mol Neurobiol 36, 1151–1159 (2016). https://doi.org/10.1007/s10571-015-0311-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10571-015-0311-6