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1.400047 重庆,重庆师范大学:教育科学学院特殊教育系
2.400047 重庆,重庆师范大学:重庆市特殊儿童心理诊断与教育技术重点实验室
3.400038 重庆,陆军军医大学(第三军医大学)基础医学院生物化学与分子生物学教研室
[ 戴旭芳,E-mail:xufangd@aliyun.com DAI Xufang, E-mail:xufangd@aliyun.com
收稿:2020-01-08,
修回:2020-03-04,
纸质出版:2020-06-30
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戴旭芳, 闫小晶, 谢鹏, 等. 丙戊酸钠通过激活miR-148a-3p/Mcl-1通路促进SH-SY5Y细胞凋亡[J]. 第三军医大学学报, 2020,42(12):1195-1200.
Xufang DAI, Xiaojing YAN, Peng XIE, et al. Sodium valproate promotes the apoptosis of SH-SY5Y cells by activating miR-148a-3p/Mcl-1 signaling pathway[J]. Journal of Third Military Medical University, 2020, 42(12): 1195-1200.
戴旭芳, 闫小晶, 谢鹏, 等. 丙戊酸钠通过激活miR-148a-3p/Mcl-1通路促进SH-SY5Y细胞凋亡[J]. 第三军医大学学报, 2020,42(12):1195-1200. DOI: 10.16016/j.1000-5404.202001055.
Xufang DAI, Xiaojing YAN, Peng XIE, et al. Sodium valproate promotes the apoptosis of SH-SY5Y cells by activating miR-148a-3p/Mcl-1 signaling pathway[J]. Journal of Third Military Medical University, 2020, 42(12): 1195-1200. DOI: 10.16016/j.1000-5404.202001055.
目的
2
探讨抗癫痫药丙戊酸钠(sodium valproate,VPA)对神经细胞的凋亡诱导作用及相关机制。
方法
2
将人神经母细胞瘤细胞(SH-SY5Y)用不同剂量VPA处理,流式分析细胞凋亡水平变化,Western blot检测Mcl-1蛋白水平变化,定量PCR分析Mcl-1 mRNA与miR-148a-3p水平变化,报告基因实验检测Mcl-1启动子活性变化。用miR-148a-3p抑制剂联合VPA处理细胞,观察miR-184a-3p在VPA下调Mcl-1表达中的作用及对VPA促SH-SY5Y细胞凋亡效果的影响。
结果
2
VPA可促进SH-SY5Y细胞凋亡,并抑制SH-SY5Y细胞中Mcl-1的蛋白和mRNA水平(
P
<
0.05)。VPA处理不影响SH-SY5Y细胞中Mcl-1的启动子活性(
P
>
0.05)。VPA可显著增强SH-SY5Y细胞中miR-148a-3p的表达(
P
<
0.05),使用miR-148a-3p抑制剂可减轻VPA对Mcl-1表达的抑制作用,并减弱VPA的促细胞凋亡效果。
结论
2
VPA可能通过激活miR-148a-3p/Mcl-1信号通路促进SH-SY5Y细胞凋亡。
Objective
2
To investigate the apoptosis-inducing effect of sodium valproate (VPA) on nerve cells and the underlying mechanism.
Methods
2
Human neuroblastoma SH-SY5Y cells treated with VPA of difterent doses were examined for apoptosis and Mcl-1 protein expression using flow cytometry and Western blotting
respectively. The mRNA expression of Mcl-1 and miR-148a-3p in the treated cells was detected with qRT-PCR
and the promoter activity of Mcl-1 gene was analyzed using a reporter gene assay. The changes of cell apoptosis and Mcl-1 expression in SH-SY5Y cells were analyzed following combined treatment with VPA and a miR-148a-3p antagonist.
Results
2
VPA treatment significantly promoted apoptosis and down-regulated Mcl-1 expression at both the mRNA and protein levels in SH-SY5Y cells. VPA treatment did not significantly affect the transcriptional activity of Mcl-1 promoter but up-regulated the expression of miR-148a-3p in SH-SY5Y cells. The application of the miR-148a-3p antagonist obviously attenuated the inhibitory effect of VPA on Mcl-1 expression and VPA-induced apoptosis in the cells.
Conclusion
2
VPA promotes the apoptosis of SH-SY5Y cells by activating miR-148a-3p/Mcl-1 signaling pathway.
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