Dement Neurocogn Disord.  2017 Dec;16(4):121-127. 10.12779/dnd.2017.16.4.121.

Neural Stem Cell Death Mechanisms Induced by Amyloid Beta

• Lee J
• Park HH
• Koh SH
• Choi H

• KMID: 2442729
• DOI: http://doi.org/10.12779/dnd.2017.16.4.121

Abstract

BACKGROUND AND PURPOSE
Amyloid beta (Aβ) is the main component of amyloid plaques, which are deposited in the brains of patients with Alzheimer's disease (AD). Biochemical and animal studies support the central role of Aβ in AD pathogenesis. Despite several investigations focused on the pathogenic mechanisms of Aβ, it is still unclear how Aβ accumulates in the central nervous system and subsequently initiates the disease at the cellular level. In this study, we investigated the pathogenic mechanisms of Aβ using proteomics and antibody microarrays.
METHODS
To evaluate the effect of Aβ on neural stem cells (NSCs), we treated primary cultured cortical NSCs with several doses of Aβ for 48 h. A 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, trypan blue staining, and bromodeoxyuridine cell proliferation assay were performed. We detected several intracellular proteins that may be associated with Aβ by proteomics and Western blotting analysis.
RESULTS
Various viability tests showed that Aβ decreased NSCs viability and cell proliferation in a concentration-dependent manner. Aβ treatment significantly decreased lactate dehydrogenase B, high-mobility group box 1, aldolase C, Ezrin, and survival signals including phosphorylated phosphoinositide 3-kinase, Akt, and glycogen synthase kinase-3β.
CONCLUSIONS
These results suggest that several factors determined by proteomics and Western blot hold the clue to Aβ pathogenesis. Further studies are required to investigate the role of these factors.

Keyword

amyloid beta; neural stem cells; proteomics

MeSH Terms

Alzheimer Disease
Amyloid*
Animals
Blotting, Western
Brain
Bromodeoxyuridine
Cell Proliferation
Central Nervous System
Fructose-Bisphosphate Aldolase
Glycogen Synthase
Humans
L-Lactate Dehydrogenase
Neural Stem Cells*
Plaque, Amyloid
Proteomics
Trypan Blue
Amyloid
Bromodeoxyuridine
Fructose-Bisphosphate Aldolase
Glycogen Synthase
L-Lactate Dehydrogenase
Trypan Blue

Figure

• 

  Fig. 1 Viability of NSCs injured by Aβ25-35. MTT assay and TBS show that the viability of NSCs was decreased in a concentration-dependent manner following treatment with more than 10 µM Aβ25-35 oligomer. *p<0.05 (vs. control group). Aβ: amyloid beta, con: control group, MTT: 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide, NSC: neural stem cell, TBS: trypan blue staining.

• 

  Fig. 2 Proliferation of NSCs injured by Aβ25-35. BrdU assay showed that the proliferation of NSCs was decreased in a concentration-dependent manner when the cells were treated with more than 10 µM Aβ25-35 oligomer. *p<0.05 (vs. control group). Aβ: amyloid beta, BrdU: bromodeoxyuridine, con: control group, NSC: neural stem cell.

• 

  Fig. 3 Effects of Aβ25-35 on intracellular protein levels of NSCs in proteomics. Proteomics showed the effects of 20 µM Aβ25-35 on various intracellular proteins in NSCs. Representative 2D-E image obtained from control group and following the treatment of 20 µM Aβ25-35 oligomer group. Protein spots with at least a two-fold difference in expression compared with the control or normal samples were selected. The proteomics results confirmed that 20 µM Aβ25-35 decreased the expression of proteins associated with cell survival. Aβ: amyloid beta, ALDOC: aldolase C, con: control group, HMGB1: high-mobility group box 1, LDHB: lactate dehydrogenase B, NSC: neural stem cell.

• 

  Fig. 4 Effects of Aβ25-35 on intracellular protein levels of NSCs in Western blot. Western blotting analysis showed that treatment with 20 µM Aβ25-35 significantly inhibited the expression of proliferation-related intracellular signaling proteins of NSCs. *p<0.05 (vs. control group). Aβ: amyloid beta, NSC: neural stem cell.

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