An Extract from Shrimp Processing By-Products Protects SH-SY5Y Cells from Neurotoxicity Induced by Aβ25–35

Abstract

Increased evidence suggests that marine unsaturated fatty acids (FAs) can protect neurons from amyloid-β (Aβ)-induced neurodegeneration. Nuclear magnetic resonance (NMR), high performance liquid chromatography (HPLC) and gas chromatography (GC) assays showed that the acetone extract 4-2A obtained from shrimp Pandalus borealis industry processing wastes contained 67.19% monounsaturated FAs and 16.84% polyunsaturated FAs. The present study evaluated the anti-oxidative and anti-inflammatory effects of 4-2A in Aβ25–35-insulted differentiated SH-SY5Y cells. Cell viability and cytotoxicity were measured by using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays. Quantitative PCR and Western blotting were used to study the expression of neurotrophins, pro-inflammatory cytokines and apoptosis-related genes. Administration of 20 μM Aβ25–35 significantly reduced SH-SY5Y cell viability, the expression of nerve growth factor (NGF) and its tyrosine kinase TrkA receptor, as well as the level of glutathione, while increased reactive oxygen species (ROS), nitric oxide, tumor necrosis factor (TNF)-α, brain derived neurotrophic factor (BDNF) and its TrkB receptor. Aβ25–35 also increased the Bax/Bcl-2 ratio and Caspase-3 expression. Treatment with 4-2A significantly attenuated the Aβ25–35-induced changes in cell viability, ROS, GSH, NGF, TrkA, TNF-α, the Bax/Bcl-2 ratio and Caspase-3, except for nitric oxide, BDNF and TrKB. In conclusion, 4-2A effectively protected SH-SY5Y cells against Aβ-induced neuronal apoptosis/death by suppressing inflammation and oxidative stress and up-regulating NGF and TrKA expression.

Inhibitory Effect of Lycopene on Amyloid-β-Induced Apoptosis in Neuronal Cells

“…It is not clear the reason why amyloid-β-induced cell death is independent on culture time. Recently, Zhang et al [ 39 ] also demonstrated that amyloid-β (20 μM)-induced cell death was higher at 24 h-culture than at 48 h-culture, which supports the present results. Further study should be performed to determine the mechanism of cell death by amyloid-β in neuronal cells using the different culture times.…”

Re-evaluation of Culture Condition of PC12 and SH-SY5Y Cells Based on Growth Rate and Amino Acid Consumption

“…We also found that treatment of amyloid peptides such as Aβ and Aβ 25-35 , nearly halved cell viability. The extent of sensitivity to amyloid peptides was similar to that of previously published values (9,11,12,13,15,18). These evidence suggests that the use of DMEM (supplemented with 10%FBS and antibiotics) is the best culture medium for both PC12 and SH-SY5Y cells.…”

“…As models of neurons, rat PC12 pheochromocytoma cells (6) and human SH-SY5Y neuroblastoma cells (7,8) have been used by many investigators to search for neuroprotective substances. However, the composition of the culture media they have used is quite different from researcher to researcher: RPMI1640 (differentiated PC12) (9, 10), RPMI (PC12) (11), DMEM (PC12) (12,13), DMEM (SH-SY5Y) (14), DMEM/Ham's F-12 (1:1) (SH-SY5Y) (15,16), MEM/F-12 (SY-SY5Y) (17) and DMEM/F-12 + non-essential amino acids (SH-SY5Y) (18), all supplemented with serum and antibiotics. It is not clear why the inclusion of F-12 and non-essential amino acids is necessary to culture SH-SY5Y cells.…”

Search of Neuroprotective Polyphenols Using the “Overlay” Isolation Method

“…However, there was no uniformity in the culture condition of PC12 and SH-SY5Y cells in previous investigations. The culture media used was: Dulbecco’s modified Eagle’s medium (DMEM); the minimum essential medium (MEM); RPMI1640; a mixture of DMEM and Ham′s F12 (1:1); and, non-essential amino acids (NEAA), which were supplemented with fetal bovine serum (FBS), alone or together with, horse serum (HS) (column A in Table 1 ) [ 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 ]. Differentiated and undifferentiated cells (column B), inoculated at different cell densities, were exposed to considerably different concentrations of neurotoxic agents (column C) in uncoated, collagen, or poly-lysine-coated plates ( Table 1 ).…”