[Effects of stachyine on apoptosis in an Aβ25-35-induced PC12 cell model of Alzheimer's disease].
Liangchao Qu, Jiajia Huang, Mingda Fan, Yuchen Hao, Jinxiu Yan
Abstract
Liangchao Qu, Jiajia Huang, Mingda Fan, Yuchen Hao, Jinxiu Yan
Abstract
OBJECTIVE To investigate the effects of stachydrine (STA) on apoptosis of Aβ25-35-induced PC12 cells mimicking Alzheimer's disease and explore the mechanisms. METHODS The differential genes of STA were analyzed based on GSE85871 data, and the target genes of STA were identified using STITCH database. PC12 cells were treated with Aβ25-35 to establish a cell model of Alzheimer's disease, and the changes in cell viability and cell cycle in response to STA treatment were assessed using MTT assay and flow cytometry, respectively. RT-PCR and Western blotting were used to detect the relevant gene or protein expressions in the treated cells. RESULTS GSE85871 data showed 37 up-regulated genes and 48 down-regulated genes in cells following treatment with STA. Analysis of the data from the STITCH database indicated that RPS8 and EED were the target genes of STA. Treatment of PC12 cells with Aβ25-35 significantly lowered the cell viability (P < 0.05) and the expressions of RPS8 and EED at both the mRNA and protein levels (P < 0.05), and obviously inhibited the expression of apoptosis-related proteins Bcl-2 and p53 (P < 0.05). STA treatment of the cells significantly reversed the effect of Aβ25-35 and induced cell cycle arrest in G2/M phase, causing also significantly increases in the expression levels of RPS8, EED, Bcl-2 and p53 (P < 0.05). CONCLUSIONS STA plays an important role in inhibiting the apoptosis of PC12 cells induced by Aβ25-35 possibly by regulating RPS8 and EED expression to promote the expressions of Bcl-2 and p53.
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OBJECTIVE To investigate the effects of stachydrine (STA) on apoptosis of Aβ25-35-induced PC12 cells mimicking Alzheimer's disease and explore the mechanisms. METHODS The differential genes of STA were analyzed based on GSE85871 data, and the target genes of STA were identified using STITCH database. PC12 cells were treated with Aβ25-35 to establish a cell model of Alzheimer's disease, and the changes in cell viability and cell cycle in response to STA treatment were assessed using MTT assay and flow cytometry, respectively. RT-PCR and Western blotting were used to detect the relevant gene or protein expressions in the treated cells. RESULTS GSE85871 data showed 37 up-regulated genes and 48 down-regulated genes in cells following treatment with STA. Analysis of the data from the STITCH database indicated that RPS8 and EED were the target genes of STA. Treatment of PC12 cells with Aβ25-35 significantly lowered the cell viability (P < 0.05) and the expressions of RPS8 and EED at both the mRNA and protein levels (P < 0.05), and obviously inhibited the expression of apoptosis-related proteins Bcl-2 and p53 (P < 0.05). STA treatment of the cells significantly reversed the effect of Aβ25-35 and induced cell cycle arrest in G2/M phase, causing also significantly increases in the expression levels of RPS8, EED, Bcl-2 and p53 (P < 0.05). CONCLUSIONS STA plays an important role in inhibiting the apoptosis of PC12 cells induced by Aβ25-35 possibly by regulating RPS8 and EED expression to promote the expressions of Bcl-2 and p53.
Key concepts: Apoptosis, Flow cytometry, Viability assay, Cell cycle, Gene, Blot, Cell, Molecular biology