南方医科大学学报

南方医科大学学报 ›› 2020, Vol. 40 ›› Issue (07): 1023-1028.doi: 10.12122/j.issn.1673-4254.2020.07.17

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水苏碱对阿尔茨海默病体外模型 Aβ25-35诱导 PC12 细胞凋亡的 影响

曲良超，黄佳佳，范明达，郝雨辰，严金秀   

  • 出版日期:2020-07-20 发布日期:2020-07-20
  • 基金资助:

Effects of stachyine on apoptosis in an Aβ25-35-induced PC12 cell model of Alzheimer’s disease

  

  • Online:2020-07-20 Published:2020-07-20

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摘要： 目的 探讨水苏碱对阿尔茨海默病体外模型Aβ25-35诱导PC12细胞凋亡的影响及其机制。方法 根据GSE85871数据分析水苏碱的差异基因，并使用STITCH数据库鉴定水苏碱的靶基因。用Aβ25-35处理的PC12细胞建立阿尔茨海默氏病体外模型，水苏碱处理PC12细胞后，MTT法测定细胞活力，通过流式细胞术检测细胞周期，并使用qRT-PCR和Western blot检测相关的基因或蛋白质的表达。结果 GSE85871数据显示，水苏碱的差异基因中有37个上调基因和48个下调基因。STITCH数据库的结果显示，RPS8和EED是水苏碱的靶基因。PC12细胞用Aβ25-35处理后，细胞的活力与对照组比较明显下降（61.06±3.32 vs 93.11±5.37，P<0.05），RPS8（0.52±0.11）和EED（0.47±0.12）基因表达水平与对照组比较明显下降下降（P<0.05），同时RPS8（0.62±0.14）和EED（0.60±0.13）蛋白表达水平与对照组（0.93±0.06）比较明显下降（P<0.05），抑制凋亡相关蛋白Bcl-2（0.36±0.02）和p53（0.35±0.03）表达降低（P<0.05）。水苏碱治疗可以改善Aβ25-35的作用，阻断G2/M期细胞周期，RPS8和EED基因表达水平明显升高（P<0.05），Bcl-2和p53的蛋白表达升高（P<0.05）。结论 水苏碱在抑制Aβ25-35诱导的PC12细胞凋亡中起重要作用，其机制可能与调节RPS8和EED从而促进Bcl-2和p53表达并抑制细胞的凋亡有关。

关键词: 水苏碱, 阿尔茨海默氏病, PC-12, 凋亡, Aβ25-35

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). Conclusion 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 words: stachydrine, Alzheimer’s disease, PC12 cells, apoptosis, Aβ25-35