Human telomerase reverse transcriptase protects human embryonic cortical neurons

doi:10.3969/j.issn.2095-4344.2013.28.021

Abstract

Abstract:

BACKGROUND:Telomerase can maintain the telomere length and avoid cell replicative senescence and apoptosis in somatic cells. Its catalytic subunit called telomerase reverse transcriptase has roles in mediating cell survival and anti-apoptotic functions.

OBJECTIVE:To evaluate the effects of human telomerase reverse transcriptase on amyloid β1-40-induced human embryonic cortical neurons injury.

METHODS: Human cortical neurons derived from 12-16 weeks old aborted fetuses were transfected with recombinant adenovirus vector encoding human telomerase reverse transcriptase. Expression of human telomerase reverse transcriptase was evaluated by immunocytochemical staining. Telomerase activity was measured using a PCR-based telomeric repeat amplification protocol enzyme-linked immunosorbent assay kit. Human embryonic cortical neurons were treated with 10 μmol/L ol/L amyloid β1-40 after transfected for 3 days. Cell viability, reactive oxygen species levels and glutathione contents in human embryonic cortical neurons were respectively detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, fluorescent probe 2',7'-dichlorodihydrofluorescein diacetate and chromatometry.

RESULTS AND CONCLUSION:Expression of human telomerase reverse transcriptase reached peak at 3 days after transfection, and the telomerase activity was rebuilt; 10 μmol/L amyloid β1-40 could significantly reduce the cell viability of neurons and glutathione content (P < 0.05 and P < 0.01), and increase the reactive oxygen species levels (P < 0.05). The neurons transfected with human telomerase reverse transcriptase gene could be significantly against the toxicity of amyloid β1-40 and increase the cell viability and glutathione content (P < 0.05 and P < 0.01), and decrease the reactive oxygen species levels (P < 0.05). The results indicate that human telomerase reverse transcriptase can protect amyloid β1-40-induced human embryonic cortical neurons injury

Key words: tissue construction, cytology experiment in tissue construction, telomerase, human telomerase reverse transcriptase, cortical neuron, β-amyloid, Alzheimer’s disease, provincial grants-supported paper

CLC Number:

R318

Wei Gui-fen, Liu Yan, Li Yan-ling, Zhang Hui-ai, Kong Ling-ping. Human telomerase reverse transcriptase protects human embryonic cortical neurons[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(28): 5229-5235.

Figures/Tables 4

References

[1]Thies W, Bleiler L. Alzheimer's Association. 2013 Alzheimer's disease facts and figures. Alzheimers Dement. 2013;9(2): 208-245.

[2]Axelsen PH, Komatsu H, Murray IV. Oxidative stress and cell membranes in the pathogenesis of Alzheimer's disease. Physiology (Bethesda). 2011;26(1):54-69.

[3]Serrano-Pozo A, Frosch MP, Masliah E, et al. Neuropathological alterations in Alzheimer disease. Cold Spring Harb Perspect Med. 2011;1(1):a006189.

[4]Galante D, Corsaro A, Florio T, et al. Differential toxicity, conformation and morphology of typical initial aggregation states of Aβ1-42 and Aβpy3-42 beta-amyloids. Int J Biochem Cell Biol. 2012;44(11):2085-2093.

[5]Zhang J, Zhen YF, Pu-Bu-Ci-Ren, et al. Salidroside attenuates beta amyloid-induced cognitive deficits via modulating oxidative stress and inflammatory mediators in rat hippocampus. Behav Brain Res. 2013;244:70-81.

[6]Hoshi A, Yamamoto T, Shimizu K, et al. Characteristics of aquaporin expression surrounding senile plaques and cerebral amyloid angiopathy in Alzheimer disease. J Neuropathol Exp Neurol. 2012;71(8):750-759.

[7]Rubtsova MP, Vasilkova DP, Malyavko AN, et al.Telomere lengthening and other functions of telomerase. Acta Nutr. 2012;4(2):44-61.

[8]Chung HK, Cheong C, Song J, et al. Extratelomeric functions of telomerase. Curr Mol Med. 2005;5(2):233-241.

[9]Niu C, Yip HK. Neuroprotective signaling mechanisms of telomerase are regulated by brain-derived neurotrophic factor in rat spinal cord motor neurons. J Neuropathol Exp Neurol. 2011;70(7):634-652.

[10]Smith JA, Park S, Krause JS, et al. Oxidative stress, DNA damage, and the telomeric complex as therapeutic targets in acute neurodegeneration. Neurochem Int. 2013;62(5):764-775.

[11]Indran IR, Hande MP, Pervaiz S. hTERT overexpression alleviates intracellular ROS production, improves mitochondrial function, and inhibits ROS-mediated apoptosis in cancer cells. Cancer Res. 2011;71(1):266-276.

[12]Fu W, Begley JG, Killen MW, et al. Anti-apoptotic role of telomerase in pheochromocytoma cells. J Biol Chem. 1999; 274(11):7264-7271.

[13]Ahmed S, Passos JF, Birket MJ, et al. Telomerase does not counteract telomere shortening but protects mitochondrial function under oxidative stress. J Cell Sci. 2008;121(Pt 7): 1046-1053.

[14]张洁,廖亚平,吴灵芝,等.hTERT基因转染对人胚胎大脑皮质神经元生长的影响[J].解剖学研究,2008,30(4):251-254.

[15]Kong LP, Wu LZ, Zhang J, et al. Neuroprotective effects of human telomerase reverse transcriptase on beta-amyloid fragment 25-35-treated human embryonic cortical neurons. Neural Regen Res. 2009;4(6):405-412.

[16]State Council of the People's Republic of China. Administrative Regulations on Medical Institution. 1994-09-01.

[17]Chandler HL, Webb TR, Barden CA, et al. The effect of phosphorylated Akt inhibition on posterior capsule opacification in an ex vivo canine model. Mol Vis. 2010;16: 2202-2214.

[18]Lustbader JW, Cirilli M, Lin C, et al. ABAD directly links Abeta to mitochondrial toxicity in Alzheimer's disease. Science. 2004; 304(5669):448-452.

[19]Casley CS, Land JM, Sharpe MA, et al. Beta-amyloid fragment 25-35 causes mitochondrial dysfunction in primary cortical neurons. Neurobiol Dis. 2002;10(3):258-267.

[20]Casley CS, Canevari L, Land JM, et al. Beta-amyloid inhibits integrated mitochondrial respiration and key enzyme activities. J Neurochem. 2002;80(1):91-100.

[21]Lamy E, Herz C, Lutz-Bonengel S, et al. The MAPK pathway signals telomerase modulation in response to isothiocyanate-induced DNA damage of human liver cancer cells. PLoS One. 2013;8(1):e53240.

[22]Zhu H, Fu W, Mattson MP. The catalytic subunit of telomerase protects neurons against Amyloid β-Peptide-Induced apoptosis. J Neurochem. 2000;75(1):117-124.

[23]Lu C, Fu W, Mattsop MP. Telomerase protects developing neurons against DNA damage–induced cell death. Brain Res Dev Brain Res. 2001;131(122):167-171.

[24]Ren JG, Xia HL, Tian YM, et al. Expression of telomerase inhibits hydroxyl radical-induced apoptosis in normal telomerase negative human lung fibroblasts. FEBS Lett. 200; 488(3):133-138.

[25]Huang XQ, Wang J, Liu JP, et al. hTERT extends proliferative lifespan and prevents oxidative stress-induced apoptosis in human lens epithelial cells. Invest Ophthalmol Vis Sci. 2005; 46(7):2503-2513.

[26]Armstrong L, Saretzki G, Peters H, et al. Overexpression of telomerase confers growth advantage, stress resistance, and enhanced differentiation of ESCs toward the hematopoietic lineage. Stem Cells. 2005;23(4):516-529.

[27]Massard C, Zermati Y, Pauleau AL, et al. hTERT: a novel endogeneous inhibitor of the mitochondrial cell death pathway. Oncogene. 2006;25(33):4505-4514.

[28]Yin L, Hubbard AK, Giardina C, et al. NF-κB regulates transcription of the mouse telomerase catalytic subunit. J Biol Chem. 200;275(47):36671-36685.

[29]Ohyagi Y, Asahara H, Chui DH, et al. Intracellular Abeta42 activates p53 promoter: a pathway to neurodegeneration in Alzheimer's disease. FASEB J. 2005;19(2):255-257.

[30]Farsetti A, Grasselli A, Bacchetti S, et al. The telomerase tale in vascular aging: regulation by estrogens and nitric oxide signaling. J Appl Physiol. 2009;106(1):333-337.

[31]Bagheri S, Nosrati M, Li S, et al. Genes and pathways downstream of telomerase in melanoma metastasis. PNAS. 2006;103(30):11306-11311.

[32]Santos JH, Meyer JN, Van Houten B. Mitochondrial localization of telomerase as a determinant for hydrogen peroxideinduced mitochondrial DNA damage and apoptosis. Hum Mol Genet. 2006;15(11):1757-1768.