Relationship between neural stem cells aging and nuclear factor erythroid 2 related factor 2/antioxidant response element signaling pathway

WU Qi; WANG Shun-He; CHENG Xiao; XIANG Yue; CHEN Lin-Bo; WANG Zi-Ling; XIAO Han-XianZhi; JIANG Rong; WANG Lu; WANG Ya-Ping

doi:10.16098/j.issn.0529.1356-2020.06.002

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Acta Anatomica Sinica ›› 2020, Vol. 51 ›› Issue (6) : 815-820. DOI: 10.16098/j.issn.0529.1356-2020.06.002

Relationship between neural stem cells aging and nuclear factor erythroid 2 related factor 2/antioxidant response element signaling pathway

WU Qi^1,2WANG Shun-he^1,2 CHENG Xiao^1,2 XIANG Yue^1,2 CHEN Lin-bo¹ WANG Zi-ling¹ XIAO Ha-xian-zhi¹ JIANG Rong¹ WANG Lu¹ WANG Ya-ping^1*

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Abstract

Objective To establish aging model of mouse neural stem cells (NSCs) in vitro and study the relationship between D-galactose (D-gal)-induced NSCs aging and nuclear factor erythroid 2 related factor 2 (Nrf2)/antioxidant response elemen (ARE) signaling pathway. Methods The whole brain of newborn C57BL/6J mouse was obtained. NSCs were isolated and identified, and cultured the cells to the third generation. The NSCs were randomly divided into control group and aging group. The NSCs of control group were cultured in NSCs medium for 48 hours, and the NSCs of aging group were added with D-galactose (D-gal,final concentration of 10 g/L) on the basis of the control group. Cell counting kit-8(CCK-8) was used to detect the activity of NSCs; Senescence-associated β-galactosidase (SA-β-gal) staining was used to detect the percentage of senescence-positive neurospheres; Trypan blue staining was used to detect cell viability; Enzyme-labeled colorimetry assay was used for detection of the activity of superoxide dismutases (SOD), catalase (CAT) and malondialdehyde (MDA) in the cell culture supernatants; Western blotting was used to detect the expression of Nrf2, heme-oxygenase-1 (HO-1) protein; Real-time PCR was applied to detect the expression level of GCLC, GCLM genes. Results Compared with the control group, the proliferative activity of NSCs in the aging group decreased significantly, the percentage of SA-β-gal staining positive neurons increased obviously, the activity of trypan blue stained NSCs decreased explicitly, SOD activity and CAT activity decreased markedly, and MDA content increased significantly. The expression levels of Nrf2/ARE signaling pathway-related proteins Nrf2 and HO-1 were significantly down-regulated in cells, the expression of pathway-associated GCLC and GCLM genes was down-regulated. Conclusion The addition of D-gal to the culture system can construct an in vitro aging model of NSCs. The possible mechanism is closely related to the inhibition of the antioxidant activity of Nrf2/ARE signaling pathway.

Key words

D-galactose / Neural stem cell / Aging / Nuclear erythroid related factor 2/antioxidant response elemen signaling pathway / Real-time PCR / Mouse

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WU Qi WANG Shun-he CHENG Xiao XIANG Yue CHEN Lin-bo WANG Zi-ling XIAO Ha-xian-zhi JIANG Rong WANG Lu WANG Ya-ping. Relationship between neural stem cells aging and nuclear factor erythroid 2 related factor 2/antioxidant response element signaling pathway[J]. Acta Anatomica Sinica. 2020, 51(6): 815-820 https://doi.org/10.16098/j.issn.0529.1356-2020.06.002

References

［1］ Ni J, Fang Y, Hu Y, et al. Effect of Tenuigenin on low density lipoprotein receptor-related protein 1 level changes of Alzheimer’s disease rat in hippocampus［J］. Acta Anatomica Sinica,2016,47(6):744-749.（in Chinese）

倪杰,房宇,胡燕,等.细叶远志皂苷对阿尔茨海默病大鼠海马低密度脂蛋白受体相关蛋白1水平的影响［J］.解剖学报,2016,47(6):744-749.

［2］ Wang X, Wang C, Pei G. α-Secretase ADAM10 physically interacts with β-secretase BACE1 in neurons and regulates CHL1 proteolysis［J］. J Mol Cell Biol, 2018, 10(5):411-422.

［3］ Dong XH, Gao WJ, Kong WN, et al. Neuroprotective effect of the active components of three Chinese herbs on brain iron load in a mouse model of Alzheimer’s disease［J］. Exp Ther Med, 2015, 9(4):1319-1327.

［4］ Zhang Y, He ML. Deferoxamine enhances alternative activation of microglia and inhibits amyloid beta deposits in APP/PS1 mice［J］. Brain Res, 2017, 1677(1):86-92.

［5］ Bilkeigorzo A. Genetic mouse models of brain ageing and Alzheimer’s disease ［J］. Pharmacol Ther, 2014, 142(2): 244-257.

［6］ Xu J, Wang J, Wimo A, et al. The economic burden of dementia in china, 1990-2030: Implications for health policy［J］. Bull World Health Organ, 2017, 95(1):18-26.

［7］ Wang ZhL, Zhang Y, Wang J. Role of Nogo-A and Nogo-A receptors in Alzheimer’s disease［J］. Acta Anatomica Sinica, 2018,49(4):549-555.（in Chinese）

王兆伦,张艳,王君. Nogo-A及其受体在阿尔茨海默病中的作用［J］.解剖学报,2018,49(4):549-555.

［8］ Dong XH, Bai JT, Kong WN, et al. Effective components of Chinese herbs reduce central nervous system function decline induced by iron overload［J］. Neural Regen Rese, 2015, 10(5): 778-785.

［9］ Dong XH, Gao WJ, Kong WN, et al. Neuroprotective effect of the active components of three Chinese herbs on brain iron load in a mouse model of Alzheimer’s disease［J］. Exp Ther Med, 2015, 9(4):1319-1327.

［10］ Ballard C, Gauthier S, Corbett A, et al. Alzheimer’s disease［J］. Lancet, 2011, 377(9770):1019-1031.

［11］ Wortmann M. World Alzheimer Report 2014:dementia and risk reduction［J］. J Alzheimer’s Assoc, 2015, 11(7): 837.

［12］ Barage SH, Sonawane KD. Amyloid cascade hypothesis: pathogenesis and therapeutic strategies in Alzheimer’s disease［J］. Neuropeptides, 2015, 52:1-18.

［13］ Endres K, Anders A, Kojro E, et al. Tumor necrosis factorD-α converting enzyme is processed by proprotein-convertases to its mature form which is degraded upon phorbol ester stimulation［J］. Eur J Biochem, 2010, 270(11):2386-2393.

［14］ Devi L, Ohno M. Effects of BACE1 haploinsufficiency on APP processing and Aβ concentrations in male and female 5XFAD Alzheimer mice at different disease stages［J］. Neuroscience, 2015, 307:128-137.

［15］ Manzine PR, Ettcheto M, Cano A, et al. ADAM10 in Alzheimer’s disease: pharmacological modulation by natural compounds and its role as a peripheral marker.［J］. Biomed Pharmacother,2019,113:108661.

［16］ Tackenberg C, Nitsch RM. The secreted APP ectodomain sAPPα, but not sAPPβ, protects neurons against Aβ oligomer-induced dendritic spine loss and increased tau phosphorylation ［J］. Mol Brain, 2019, 12(1):27.

［17］ Peterslibeu C, Campagna J, Mitsumori M, et al. sAβPPα is a potent endogenous inhibitor of BACE1［J］. J Alzheimers Dis, 2015, 47(3):545-555.

［18］ Barger SW, Harmon AD. Microglial activation by Alzheimer amyloid precursor protein and modulation by apolipoprotein E ［J］. Nature, 1998, 388(6645):878-881.

［19］ Su X, Zhang Zh, Yang LJ, et al. Granulocyte colony stimulating factor improving the earning and memory function and its anti-inflammatory mechanism of Alzheimer’s disease model rats［J］. Acta Anatomica Sinica, 2019,50(1):29-34.（in Chinese）

苏翔,张臻,杨霖璟,等.粒细胞集落刺激因子改善阿尔茨海默病模型大鼠学习记忆功能及抗炎作用机制［J］.解剖学报,2019,50(1):29-34.

［20］ Joseph JA, Carey A, Brewer GJ, et al. Dopamine and a beta-induced stress signaling and decrements in Ca2+ buffering in primary neonatal hippocampal cells are antagonized by blueberry extract［J］. J Alzheimers Dis, 2007, 11(4):433-446.

［21］ Luo XJ, Lin M, Wang ChM. Role and research progress of inflammation in the mechanism of Alzheimer’s disease［J］.Journal of Tropical Medicine, 2016(1):119-122.（in Chinese）

罗小金, 林茂, 王春梅. 炎症反应在阿尔茨海默病发病机制中的研究进展［J］. 热带医学杂志, 2016(1):119-122.