Study on the toxic effects of Aβ1-42 oligomers in SHSY5Y with human A53T mutant α-synuclein over-expression.
10.3969/j.issn.1002-0152.2019.07.008
- VernacularTitle:Aβ1-42寡聚体对α-syn过表达SHSY5YA53T细胞自噬功能的影响☆
- Author:
Manli GUO
1
,
2
,
3
;
Yuyuan GAO
;
Qingxi ZHANG
;
Kun NIE
;
Lijuan. WANG
Author Information
1. 华南理工大学医学院(广州510006)
2. 广东省人民医院(广东省医学科学院)神经科
3. 广东省神经科学研究所
- Keywords:
Alpha-synuclein;
Beta-amyloid 1-42 oligomer;
Parkinson disease;
Autophagy
- From:
Chinese Journal of Nervous and Mental Diseases
2019;45(7):422-426
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish an in vitro cell model of Parkinson disease with SHSY5Y cells over-expressing human A53T mutant alpha-synuclein and to examine the effects of Aβ1-42 oligomer on cell survival and autophagy function in the cell model Method The recombinant lentivirus containing the A53T mutant alpha-synuclein gene or empty vector were transfected to SHSY5Y cells. The expression of α-synuclein mRNA in SHSY5Y cells was detected by RT-qPCR. The effect of Aβ1-42 oligomer on cell proliferation was detected with CCK-8 after incubation with Aβ1-42 oligomer for 24 hours. The autophagy-related proteins were evaluated with Western Blot. Result The mRNA and protein levels of alpha-synuclein were significantly increased in SHSY5Y cells expressing alpha-synuclein. There were no significant difference in the cell proliferation between alpha-synuclein group and control group (P<0.001) . Incubation with Aβ1-42 oligomer significantly decreased the proliferation rate in alpha-synuclein group in a dose-dependent manner compared with the control group. The levels of autophagy related proteins including LC3-Ⅱ and Beclin-1 were significantly lower in alpha-synuclein group than in control group (P<0.05). Conclusion This work has constructed an in vitro cell model of Parkinson′s disease. The over-expression of A53T mutant alpha-synuclein do not affect the cell survival whereas the Aβ1-42 oligomer exhibits toxic effects on cells expressing alpha-synuclein possible through suppression of the autophagy activation.
