依达拉奉通过microRNA-25抑制高糖诱导的SH-SY5Y细胞凋亡

发布时间：2018-06-06 05:18

  本文选题：依达拉奉 + 微小RNA-　； 参考：《中国病理生理杂志》2017年01期

【摘要】：目的:探讨依达拉奉通过微小RNA-25(microRNA-25,miR-25)对高糖诱导的人神经母细胞瘤SHSY5Y细胞凋亡的抑制作用及其机制。方法:将SH-SY5Y细胞用含高浓度葡萄糖的DMEM培养基和依达拉奉的联合培养液共同培养24 h。MTT比色法测定SH-SY5Y细胞存活率;DCFH-DA荧光探针法检测SH-SY5Y细胞中活性氧簇(ROS)的水平;采用流式细胞术检测SH-SY5Y细胞的凋亡率;Western blot法检测凋亡相关蛋白Bax和Bcl-2的表达水平;实时定量PCR检测细胞中miR-25的表达水平。为进一步阐明依达拉奉抑制高糖诱导的神经细胞凋亡的作用靶点,我们将miR-25抑制剂应用于细胞,之后采用caspase-3凋亡试剂盒检测细胞的凋亡率。结果:与对照组相比,高糖诱导后细胞存活率明显降低,细胞中的ROS水平和细胞凋亡率明显升高,Bax的表达明显增加,Bcl-2的表达明显降低,miR-25的表达水平也明显降低。给予依达拉奉治疗之后,细胞存活率明显升高,ROS含量和细胞凋亡率明显降低,Bax的蛋白水平明显降低,Bcl-2蛋白水平明显升高,miR-25的表达水平亦明显升高。进一步给予miR-25抑制剂后,caspase-3的水平明显升高,此时同时给予依达拉奉后并不能抑制高糖引起的神经细胞的凋亡。结论:依达拉奉对高糖诱导的SH-SY5Y细胞凋亡具有抑制作用,其作用靶点可能是miR-25。
[Abstract]:Aim: to investigate the inhibitory effect of Edaravone on the apoptosis of human neuroblastoma SHSY5Y cells induced by high glucose and its mechanism. Methods: SH-SY5Y cells were cultured in DMEM medium with high concentration of glucose and Edaravone co-cultured in 24 h.MTT colorimetric assay. The survival rate of SH-SY5Y cells was determined by fluorescence probe method. The level of Ros in SH-SY5Y cells was detected by fluorescence probe method. The apoptotic rate of SH-SY5Y cells was detected by flow cytometry and the expression of Bax and Bcl-2 was detected by Western blot, and the expression of miR-25 was detected by real-time quantitative PCR. To further elucidate the role of Edaravone in inhibiting the apoptosis of neurons induced by high glucose, we applied the miR-25 inhibitor to the cells, and then used the caspase-3 apoptosis kit to detect the apoptosis rate of the cells. Results: compared with the control group, the survival rate of the cells induced by high glucose was significantly decreased, the ROS level and the apoptosis rate of the cells were significantly increased, the expression of Bcl 2 was significantly increased and the expression level of miR-25 was significantly decreased. After treatment with Edaravone, the cell survival rate and apoptosis rate were significantly increased, the protein level of Bax was significantly decreased, the level of Bcl 2 protein was significantly increased, the expression level of miR-25 was significantly increased. The level of caspase-3 increased significantly after further administration of miR-25 inhibitor, but Edaravone did not inhibit the neuronal apoptosis induced by high glucose. Conclusion: Edaravone can inhibit the apoptosis of SH-SY5Y cells induced by high glucose, and its target may be miR-25.
【作者单位】： 中国人民解放军第四零一医院急诊科;滨州医学院附属医院老年内科;
【分类号】：R587.1
，

本文编号：1985291