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[Establishment of a new cell model mimicking Alzheimer's disease by knocking down expression].

作者信息

Luo Jing, Zhao Yan, Xie Jingwen, Liu Xin, Lin Fangbo, Hou Deren

机构信息

Department of Neurology, Third Xiangya Hospital of Central South University, Changsha 410013, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2018 Jan 30;38(1):8-13. doi: 10.3969/j.issn.1673-4254.2018.01.02.

Abstract

OBJECTIVE

To establish a cell model mimicking Alzheimer's disease (AD) by knocking down gene and compare the viability, apoptosis, and expressions of tumor necrosis factor- (TNF-) 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 -shRNA lentiviral vector, and 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- and IL -1β levels 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 ( < 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 (>0.05). Significantly increased expressions of TNF- and IL-1β were observed in both of the two cell models compared with their respective control groups ( < 0.05) without significant differences between the two cell models or between the two control groups (>0.05).

CONCLUSIONS

A new AD cell model similar to Aβ25-35-induced AD model can be established by knockdown in N2a cells.

摘要

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