Anti-autophagic and anti-apoptotic effects of memantine in a SH-SY5Y cell model of Alzheimer's disease via mammalian target of rapamycin-dependent and -independent pathways

Song,Guijun; Li,Yu; Lin,Lulu; Cao,Yunpeng

doi:10.3892/mmr.2015.4382

Anti-autophagic and anti-apoptotic effects of memantine in a SH-SY5Y cell model of Alzheimer's disease via mammalian target of rapamycin-dependent and -independent pathways

Authors:
- Guijun Song
- Yu Li
- Lulu Lin
- Yunpeng Cao
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Affiliations: Department of Neurology, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China
Published online on: September 29, 2015 https://doi.org/10.3892/mmr.2015.4382
Pages: 7615-7622
Copyright: © Song et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Memantine non-competitively blocks the N-methyl-D-aspartate receptor in order to inhibit beta-amyloid (Aβ) secretion, and has been used to treat moderate-to-severe Alzheimer's disease (AD). However, the mechanisms underlying the role of memantine in the autophagy and apoptosis of neuronal cells in AD, as well as the association between neuronal autophagy and apoptosis have yet to be elucidated. The present study aimed to establish an AD cell model overexpressing the 695‑amino‑acid Swedish mutant of Aβ precursor protein (APP695swe) in order to observe the effects of memantine on the cell viability, autophagy and apoptosis of SH‑SY5Y cells in the AD model, and to investigate the associated underlying mechanisms. A pcDNA3.1‑APP695 plasmid was transfected into the SH‑SY5Y cells. Reverse transcription‑quantitative polymerase chain reaction and western blot analyses demonstrated that the AD cell model was successfully established. MTT assays demonstrated that memantine was able to upregulate neuronal cell survival, and acridine orange staining and flow cytometry demonstrated that memantine (5 µM) was able to inhibit neuronal autophagy and apoptosis. Following neuronal autophagy induction by rapamycin, cell apoptosis rates increased significantly. Further experiments revealed that memantine was able to upregulate the expression of signaling molecules phosphorylated (p)-phosphoinositide 3‑kinase, p‑Akt and p‑mammalian target of rapamycin (mTOR), and also inhibited the phosphorylation of the B‑cell lymphoma 2/Beclin‑1 complex via mitogen‑activated protein kinase 8. In conclusion, the results of the present study demonstrated that in the AD cell model, autophagy was able to promote apoptosis. Memantine exerted anti‑autophagic and anti‑apoptotic functions, and mTOR‑dependent as well as ‑independent autophagic signaling pathways were involved in this process. Therefore, these results of the present study strongly supported the use of memantine as a potential therapeutic strategy for AD treatment.

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1	Ballard C, Gauthier S, Corbett A, Brayne C, Aarsland D and Jones E: Alzheimer's disease. Lancet. 377:1019–1031. 2011. View Article : Google Scholar : PubMed/NCBI
2	Huang Y and Mucke L: Alzheimer mechanisms and therapeutic strategies. Cell. 148:1204–1222. 2012. View Article : Google Scholar : PubMed/NCBI
3	De Felice FG, Velasco PT, Lambert MP, Viola K, Fernandez SJ, Ferreira ST and Klein WL: Abeta oligomers induce neuronal oxidative stress through an N- methyl-d- aspartate receptor- dependent mechanism that is blocked by the Alzheimer drug memantine. J Biol Chem. 282:11590–11601. 2007. View Article : Google Scholar : PubMed/NCBI
4	Texidó L, Martín-Satué M, Alberdi E, Solsona C and Matute C: Amyloid β peptide oligomers directly activate NMDA receptors. Cell Calcium. 49:184–190. 2011. View Article : Google Scholar
5	Zheng L, Kågedal K, Dehvari N, Benedikz E, Cowburn R, Marcusson J and Terman A: Oxidative stress induces macroautophagy of amyloid beta- protein and ensuing apoptosis. Free Radic Biol Med. 46:422–429. 2009. View Article : Google Scholar
6	Lorenzi M, Beltramello A, Mercuri NB, Mercuri NB, Canu E, Zoccatelli G, Pizzini FB, Alessandrini F, Cotelli M and Rosini S: Effect of memantine on resting state default mode network activity in Alzheimer's disease. Drugs Aging. 28:205–217. 2011. View Article : Google Scholar : PubMed/NCBI
7	Saxton J, Hofbauer RK, Woodward M, Gilchrist NL, Potocnik F, Hsu HA, Miller ML, Pejović V, Graham SM and Perhach JL: Memantine and functional communication in Alzheimer's disease: Results of a 12- week, international, randomized clinical trial. J Alzheimers Dis. 28:109–118. 2012.
8	Schulz JB, Rainer M, Klünemann HH, Kurz A, Wolf S, Sternberg K and Tennigkeit F: Sustained effects of once - daily memantine treatment on cognition and functional communication skills in patients with moderate to severe Alzheimer's disease: Results of a 16- week open- label trial. J Alzheimers Dis. 25:463–475. 2011.
9	Liu MY, Wang S, Yao WF, Zhang ZJ, Zhong X, Sha L, He M, Zheng ZH and Wei MJ: Memantine improves spatial learning and memory impairments by regulating NGF signaling in APP/PS1 transgenic mice. Neuroscience. 273:141–151. 2014. View Article : Google Scholar : PubMed/NCBI
10	Danysz W and Parsons CG: The NMDA receptor antagonist memantine as a symptomatological and neuroprotective treatment for Alzheimer's disease: Preclinical evidence. Int J Geriatr Psychiatry (Suppl 1). 18:S23–S32. 2003. View Article : Google Scholar
11	Peskind ER, Potkin SG, Pomara N, Ott BR, Graham SM, Olin JT and McDonald S: Memantine treatment in mild to moderate Alzheimer disease: A 24- week randomized, controlled trial. Am J Geriatr Psychiatry. 14:704–715. 2006. View Article : Google Scholar : PubMed/NCBI
12	Ray B, Banerjee PK, Greig NH and Lahiri DK: Memantine treatment decreases levels of secreted Alzheimer's amyloid precursor protein (APP) and amyloid beta (A beta) peptide in the human neuroblastoma cells. Neurosci Lett. 470:1–5. 2010. View Article : Google Scholar
13	Alley GM, Bailey JA, Chen D, Ray B, Puli LK, Tanila H, Banerjee PK and Lahiri DK: Memantine lowers amyloid- beta peptide levels in neuronal cultures and in APP/PS1 transgenic mice. J Neurosci Res. 88:143–154. 2010. View Article : Google Scholar
14	Maekawa M, Namba T, Suzuki E, Yuasa S, Kohsaka S and Uchino S: NMDA receptor antagonist memantine promotes cell proliferation and production of mature granule neurons in the adult hippocampus. Neurosci Res. 63:259–266. 2009. View Article : Google Scholar : PubMed/NCBI
15	Namba T, Maekawa M, Yuasa S, Kohsaka S and Uchino S: The Alzheimer's disease drug memantine increases the number of radial glia-like progenitor cells in adult hippocampus. Glia. 57:1082–1090. 2009. View Article : Google Scholar
16	Namba T, Yabe T, Gonda Y, et al: Pigment epithelium-derived factor upregulation induced by memantine, an N-methyl-d-aspartate receptor antagonist, is involved in increased proliferation of hippocampal progenitor cells. Neuroscience. 167:372–383. 2010. View Article : Google Scholar : PubMed/NCBI
17	Ghavami S, Shojaei S, Yeganeh B, Ande SR, Jangamreddy JR, Mehrpour M, Christoffersson J, Chaabane W, Moghadam AR and Kashani HH: Autophagy and apoptosis dysfunction in neuro-degenerative disorders. Prog Neurobiol. 112:24–49. 2014. View Article : Google Scholar
18	Nixon RA: Autophagy, amyloidogenesis and Alzheimer disease. J Cell Sci. 120:4081–4091. 2007. View Article : Google Scholar : PubMed/NCBI
19	Nilsson P, Loganathan K, Sekiguchi M, Matsuba Y, Hui K, Tsubuki S, Tanaka M, Iwata N, Saito T and Saido TC: Aβ secretion and plaque formation depend on autophagy. Cell Reports. 5:61–69. 2013. View Article : Google Scholar
20	Yu WH, Cuervo AM, Kumar A, Peterhoff CM, Schmidt SD, Lee JH, Mohan PS, Mercken M, Farmery MR and Tjernberg LO: Macroautophagy– a novel Beta- amyloid peptide- generating pathway activated in Alzheimer's disease. J Cell Biol. 171:87–98. 2005. View Article : Google Scholar : PubMed/NCBI
21	Spilman P, Podlutskaya N, Hart MJ, Debnath J, Gorostiza O, Bredesen D, Richardson A, Strong R and Galvan V: Inhibition of mTOR by rapamycin abolishes cognitive deficits and reduces amyloid- beta levels in a mouse model of Alzheimer's disease. PLoS One. 5:e99792010. View Article : Google Scholar
22	Caccamo A, Majumder S, Richardson A, Strong R and Oddo S: Molecular interplay between mammalian target of rapamycin (mTOR), amyloid- beta, and Tau: Effects on cognitive impairments. J Biol Chem. 285:13107–13120. 2010. View Article : Google Scholar : PubMed/NCBI
23	Tan CC, Yu JT, Tan MS, Jiang T, Zhu XC and Tan L: Autophagy in aging and neurodegenerative diseases: Implications for pathogenesis and therapy. Neurobiol Aging. 35:941–957. 2014. View Article : Google Scholar
24	Zhu XC, Yu JT, Jiang T and Tan L: Autophagy modulation for Alzheimer's disease therapy. Mol Neurobiol. 48. pp. 702–714. 2013, View Article : Google Scholar
25	Ye X, Tai W, Bao X, Chen X and Zhang D: FLZ inhibited γ- secretase selectively and decreased Aβ mitochondrial production in APP- SH- SY5Y cells. Naunyn Schmiedebergs Arch Pharmacol. 387:75–85. 2014. View Article : Google Scholar
26	Shen YE, Wang Y, Yu GC, Liu C, Zhang ZY and Zhang LM: Effects of edaravone on amyloid-β precursor protein processing in SY5Y- APP695 cells. Neurotox Res. 24:139–147. 2013. View Article : Google Scholar : PubMed/NCBI
27	Wang L, Zhang D, Yu Y, Guan H, Qiao C and Shang T: RNA interference- mediated silencing of laminin receptor 1 (LR1) suppresses migration and invasion and downregulates matrix metalloproteinase (MMP)- 2 and MMP- 9 in trophoblast cells: Implication in the pathogenesis of preeclampsia. J Mol Histol. 44:661–668. 2013. View Article : Google Scholar : PubMed/NCBI
28	Nixon RA, Wegiel J, Kumar A, Yu WH, Peterhoff C, Cataldo A and Cuervo AM: Extensive involvement of autophagy in Alzheimer disease: An immunoelectron microscopy study. J Neuropathol Exp Neurol. 64:113–122. 2005.PubMed/NCBI
29	Louneva N, Cohen JW, Han LY, albot K, Wilson RS, Bennett DA, Trojanowski JQ and Arnold SE: Caspase- 3 is enriched in post-synaptic densities and increased in Alzheimer's disease. Am J Pathol. 173:1488–1495. 2008. View Article : Google Scholar : PubMed/NCBI
30	Kaufman AM, Milnerwood AJ, Sepers MD, Coquinco A, She K, Wang L, Lee H, Craig AM, Cynader M and Raymond LA: Opposing roles of synaptic and extrasynaptic NMDA receptor signaling in cocultured striatal and cortical neurons. J Neurosci. 32:3992–4003. 2012. View Article : Google Scholar : PubMed/NCBI
31	Caballero B and Coto- Montes A: An insight into the role of autophagy in cell responses in the aging and neurodegenerative brain. Histol Histopathol. 27:263–275. 2012.PubMed/NCBI
32	Gao X, Xu X, Pang J, Zhang C, Ding JM, Peng X, Liu Y and Cao JM: NMDA receptor activation induces mitochondrial dysfunction, oxidative stress and apoptosis in cultured neonatal rat cardiomyocytes. Physiol Res. 56:559–569. 2007.
33	Han W, Pan H, Chen Y, Sun J, Wang Y, Li J, Ge W, Feng L, Lin X, Wang X, et al: EGFR tyrosine kinase inhibitors activate autophagy as a cytoprotective response in human lung cancer cells. PLoS One. 6:e186912011. View Article : Google Scholar : PubMed/NCBI
34	Liu J, Hu XJ, Jin B, Qu XJ, Hou KZ and Liu YP: β- Elemene induces apoptosis as well as protective autophagy in human non- small- cell lung cancer A549 cells. J Pharm Pharmacol. 64:146–153. 2012. View Article : Google Scholar
35	Wang P, Guo QS, Wang ZW and Qian HX: HBx induces HepG- 2 cells autophagy through PI3K/Akt- mTOR pathway. Mol Cell Biochem. 372:161–168. 2013. View Article : Google Scholar
36	Yang YH, Chen K, Li B, Chen JW, Zheng XF, Wang YR, Jiang SD and Jiang LS: Estradiol inhibits osteoblast apoptosis via promotion of autophagy through the ER- ERK- mTOR pathway. Apoptosis. 18:1363–1375. 2013. View Article : Google Scholar : PubMed/NCBI
37	Russell RC, Tian Y, Yuan H, Park HW, Chang YY, Kim J, Kim H, Neufeld TP, Dillin A and Guan KL: ULK1 induces autophagy by phosphorylating Beclin- 1 and activating VPS34 lipid kinase. Nat Cell Biol. 15:741–750. 2013. View Article : Google Scholar : PubMed/NCBI