丙戊酸钠通过激活miR-148a-3p/Mcl-1通路促进SH-SY5Y细胞凋亡

Sodium valproate promotes the apoptosis of SH-SY5Y cells by activating miR-148a-3p/Mcl-1 signaling pathway

目的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 To investigate the apoptosis-inducing effect of sodium valproate(VPA)on nerve cells and the underlying mechanism.Methods 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 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 VPA promotes the apoptosis of SH-SY5Y cells by activating miR-148a-3p/Mcl-1 signaling pathway.