摘要
目的 应用体外模型诱导人神经母细胞瘤SH-SY5Y细胞氧糖剥夺(OGD),探讨神经元在缺血性损伤中的潜在机制。方法 应用糖氧剥夺模型及SH-SY5Y细胞进行OGD处理,时间设为0h、12h及24h,应用活性氧(ROS)检测试剂盒检测各组细胞中ROS水平;使用MTT法检测各组细胞的生存率;WesternBlotting检测各组细胞中凋亡诱导因子(AIF)蛋白、多聚ADP核糖聚合酶-1(PARP1)蛋白及PAR蛋白的表达变化;使用线粒体膜电位检测试剂盒(JC-1)检测细胞中线粒体内膜电位变化。结果 随着OGD时间的延长,MTT检测SH-SY5Y细胞的生存率逐渐降低(P<0.05);WesternBlotting检测显示OGD明显增加了细胞中AIF蛋白、PARP1蛋白及PAR蛋白的表达水平(P<0.05);JC-1检测显示随着OGD作用时间的延长,线粒体膜电位逐渐降低;在使用抗氧化抑制剂N-乙酰半胱氨酸(NEC)后,SH-SY5Y细胞中AIF蛋白、PARP1蛋白及PAR蛋白表达量明显降低,且细胞生存率也显著提高(P<0.05)。结论 氧糖剥夺通过上调ROS水平介导SH-SY5Y细胞Parthanatos死亡。
Objective By inducing oxygen-glucose deprivation(OGD)in human neuroblastoma SH-SY5Y cells using an in vitro model to explore the potential mechanism of neurons in ischemic injury. Methods Oxygen deprivation model and SH-SY5Y cells were used for OGD treatment.The time was set to 0 hours,12 hours and 24 hours.Reactive oxygen species(ROS)were detected by reactive oxygen species detection kit.Cell viability was detected by MTT.The expression of AIF protein,PARP1 protein and PAR protein in each group was detected by Western Blotting.The mitochondrial membrane potential was detected by mitochondrial membrane potential detection kit(JC-1). Results With the prolongation of OGD time,the survival rate of SH-SY5Y cells decreased gradually by MTT( P <0.05).Western Blotting showed that OGD significantly increased the protein expression levels of AIF,PARP1 and PAR in cells;JC-1 detection shown that the mitochondrial membrane potential gradually decreased with the prolongation of OGD action time.After using the antioxidant inhibitor N-acetylcysteine(NEC),the expression of AIF,PARP1 and PAR in SH-SY5Y cells was significantly decreased.And the cell survival rate was also significantly increased( P <0.05). Conclusion Oxygen glucose deprivation mediates the death of parthanatos in SH-SY5Y cells by up-regulating ROS levels.
作者
郭文旭
张忠敏
关亚新
GUO Wen-xu(Department of Neurology,Hongqi Hospital,Affiliated to Mudanjiang Medical College,Mudanjiang 157011,China)
出处
《牡丹江医学院学报》
2019年第3期1-4,共4页
Journal of Mudanjiang Medical University
