RUNX1对氧糖剥夺诱导PC12细胞凋亡的保护作用研究

目的:研究RUNX1在PC12细胞氧糖剥夺模型中的表达及其对PC12细胞的保护作用,并探讨其相关机制。方法:体外培养PC12细胞并构建氧糖剥夺模型,将细胞分为对照组、氧糖剥夺组、RUNX1 si RNA处理组、si RNA对照处理组(sicontrol)、pc DNA3.1-RUNX1处理组(pc RUNX1)和pc DNA3.1对照处理组(pc DNA 3.1)。q RT-PCR和western blot检测RUNX1、磷酸化Akt(p-Akt)和总Akt(t-Akt)表达水平;MTT法检测细胞存活率;Annexin V-FITC/PI双染法检测细胞凋亡。结果:与对照组比较,RUNX1在PC12细胞氧糖剥夺模型中表达水平显著升高;沉默RUNX1可下调PC12细胞的存活率,促进细胞的凋亡,有效抑制p-Akt蛋白表达,而过表达RUNX1显著提高细胞存活率,抑制细胞凋亡,并上调p-Akt蛋白表达;此外,PI3K/Akt通路抑制剂LY294002明显抑制RUNX1过表达对细胞存活率的促进作用和对细胞凋亡的抑制作用。结论:RUNX1可通过PI3K/Akt信号通路保护OGD对PC12细胞的损伤作用。

Protective Effects of RUNX1 on Oxygen-glucose Deprivation Induced Apoptosis in PC12 Cells

ABSTRACT Objective: To estimate the expression of RUNX1 in oxygen-glucose deprivation injured PC12 cells and investigate the protective effect and mechanisms of RUNX1 on apoptosis induced by oxygen-glucose deprivation in PC12 cells. Methods: The oxygen-glucose deprivation (OGD) model was built using PC12 cells, and cells were divided into control group, OGD group, RUNX1 siRNA group, control siRNA group, pcDNA3.1-RUNX1 group and pcDNA3.1 group. The expression of RUNX1, phosphorylated Akt (p-Akt) and total Akt (t-Akt) were measured using qRT-PCR and western blot assay. Cell viability was estimated using MTT assay and cell apoptosis was detected using Annexin V-FITC/PI assay. Results: Compared with control, RUNX1 expression was significantly higher in OGD model. Silence RUNX1 by RUNX1 siRNA remarkably inhibited cell viability, promoted apoptosis and down-regulated p-Akt protein expression, while RUNX1 overexpression showed a contrary effect. Besides, LY2904002, the inhibitor of PI3K/Akt, markedly impeded the the role of RUNX1 overexpression in promoting cell viability and inhibiting apoptosis. Conclusion: RUNX1 protected PC12 cells against oxygen-glucose deprivation injury through PI3K/Akt signaling pathway.