[Establishment of a New Cell Model Mimicking Alzheimer's Disease by Knocking Down SORL1 Expression].
Nan fang yi ke da xue xue bao = Journal of Southern Medical University(2018)
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-α 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 (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-α and IL-1β were 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). CONCLUSIONS:A new AD cell model similar to Aβ25-35-induced AD model can be established by SORL1 knockdown in N2a cells.
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Key words
SORL1,Alzheimer's disease,N2a cell,transfection
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