芍药苷对PINK1-Parkin介导的线粒体自噬在H2O2诱导SH-SY5Y细胞损伤中的影响

Effects of Paeoniflorin on PINK1-Parkin Mediated Mitophagy in H2O2-induced SH-SY5Y Cell Damage

目的观察芍药苷对过氧化氢(H2O2)诱导的人神经母细胞瘤细胞(SH-SY5Y)细胞损伤中线粒体自噬的影响,探讨其相关作用机制。方法采用250μmol/L H2O2诱导SH-SY5Y细胞24 h构建细胞损伤模型。实验细胞分为空白组、模型组和芍药苷低、中、高剂量组。采用MTT法检测细胞存活率,JC-1线粒体膜电位荧光探针检测线粒体膜电位,TUNEL染色检测细胞凋亡率,Western blot检测PINK1、Parkin、LC3Ⅱ和p62蛋白的表达。结果与空白组比较,模型组细胞存活率下降,线粒体膜电位下降,细胞凋亡率上升,PINK1、LC3Ⅱ和p62蛋白表达升高,Parkin蛋白表达无明显变化;与模型组比较,芍药苷低、中、高剂量组细胞线粒体膜电位上调,细胞凋亡率下降,PINK1、LC3Ⅱ和p62蛋白表达降低,Parkin蛋白表达升高。结论芍药苷可显著降低H2O2诱导的SH-SY5Y细胞损伤,其机制可能与调控PINK1-Parkin介导的线粒体自噬途径相关。

Objective To explore the regulatory effects of paeoniflorin on mitophagy in H2O2-induced SH-SY5Y cell damage;To discuss its mechanism of action.Methods 250μmol/L H2O2 was used to induce SH-SY5Y for 24 h to construct a cell injury model.The blank group,model group and paeoniflorin low-,medium-and high-dosage groups were involved in the experiment.Methyl thiazolyl tetrazolium assay was used to detect cell survival rate.JC-1 mitochondrial membrane potential fluorescent probe was used to detect mitochondrial membrane potential.TUNEL staining was used to detect apoptosis rate.Western blot was adopted to detect the protein expression levels of PINK1,Parkin,LC3Ⅱand p62.Results Compared with the blank group,the cell survival rate of the model group decreased;the mitochondrial membrane potential decreased;the apoptosis rate increased;the expressions of PINK1,LC3Ⅱand p62 protein increased;the expression of Parkin protein did not change significantly.Compared with model group,the mitochondrial membrane potential of paeoniflorin low-,medium-and high-dosage groups increased;the apoptosis rate decreased;the expressions of PINK1,LC3Ⅱand p62 protein decreased;the expression of Parkin protein increased.Conclusion Paeoniflorin can significantly reduce H2O2-induced SH-SY5Y cell damage,and the mechanism of action may be related to the regulation of PINK1-Parkin-mediated mitophagy pathway.