Establishment of a new cell model mimicking Alzheimer's disease by knocking down SORL1 expression
10.3969/j.issn.1673-4254.2018.01.002
- VernacularTitle:敲低SORL1表达构建模拟阿尔茨海默病的细胞模型
- Author:
Jing LUO
1
;
Yan ZHAO
;
Jingwen XIE
;
Xin LIU
;
Fangbo LIN
;
Deren HOU
Author Information
1. 中南大学湘雅三医院神经内科
- Keywords:
SORL1;
Alzheimer's disease;
N2a cell;
transfection
- From:
Journal of Southern Medical University
2018;38(1):8-13
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a cell model mimicking Alzheimer's disease (AD) by knocking down SORL1 gene and compare the viability, apoptosis, and expressions of tumor necrosis factor-α( TNF-α) and interleukin-1β(IL-1β) in this model with a traditional Alzheimer's disease cell model. Methods A traditional cell model of AD was established by inducing N2a cells with Aβ25-35, and the optimal Aβ25-35 concentration was determined by assessing the cell viability changes. Another cell model of AD was established by transfecting N2a cells with SORL1-shRNA lentiviral vector, and SORL1 expression in the transfected cells were detected using Western blotting and qRT-PCR. With wild-type N2a cells without any treatment and cells transfected with a scramble shRNA as the control groups, the two cell models were examined for cell viability with MTT assay, cell apoptosis with flow cytometry, and TNF-αand IL-1βlevels in the culture supernatant with ELISA. Results The two cell models of AD showed obviously decreased viability and increased cell apoptosis compared with the untreated control cells or cells transfected with a scramble shRNA (P<0.05); no significant difference was found in the cell viability and apoptosis rate between the two AD cell models or between the two control groups (P>0.05). Significantly increased expressions of TNF-αand IL-1βwere observed in both of the two cell models compared with their respective control groups (P<0.05) without significant differences between the two cell models or between the two control groups (P>0.05). Conclusion A new AD cell model similar to Aβ25-35-induced AD model can be established by SORL1 knockdown in N2a cells.
