IGF-1对6-OHDA诱导神经元氧化损伤的保护作用

The protective effect of IGF-1 on 6-OHDA induced oxidative damage in neurons

目的探讨胰岛素样生长因子-1(IGF-1)对6-羟基多巴胺(6-OHDA)诱导的PC12细胞氧化损伤的保护作用。方法以25、50、100、150、200μmol/L的6-OHDA处理PC12细胞,在12、24、48 h用MTT法检测6-OHDA对PC12细胞活性的影响,筛选最佳的实验浓度和观察时间。实验分为3组:对照组、6-OHDA组(150μmol/L处理24 h)和IGF-1+6-OHDA组(IGF-1 100 nmol/L预处理2 h,后加入150μmol/L 6-OHDA处理24 h),MTT法检测各组细胞活性;免疫荧光染色法检测细胞活性氧(ROS)水平;Hoechst33342/PI双染法检测细胞凋亡。结果随6-OHDA浓度的增加和作用时间的延长,PC12细胞的活性呈梯度降低,150μmol/L 6-OHDA浓度和处理后24 h作为本研究的最佳的实验浓度和观察时间。与6-OHDA组比较,IGF-1+6-OHDA组PC12细胞活力增强、ROS水平下降、细胞凋亡减少。结论 IGF-1预处理能减少6-OHDA引起的PC12细胞氧化损伤及凋亡,增加细胞活性,为防治帕金森病提供了潜在的治疗策略。

Objective To investigate the protective effect of insulin-like growth factor-1(IGF-1)on 6-hydroxy dopamine(6-OHDA)induced oxidative damage in PC-12 cells.Methods PC12 cells were treated with 6-OHDA(concentrations of 25,50,100,150 and 200μmol/L).In order to select the optimal experimental concentration and treatment time,the activity of PC12 cells was detected by MTT at different time points of 12 h,24 h and 48 h after treatment.PC12 cells were divided into three groups:control group,6-OHDA group and IGF-1+6-OHDA group.The activity of PC12 cells was detected by MTT assay.Reactive oxygen species(ROS)level of PC12 cells was detected by immunofluorescence staining,and apoptosis was detected by Hoechst33342/PI double staining method.Results With the increased concentration and the prolongation of the action time of 6-OHDA,the activity of PC12 cells decreased gradually.The concentration of 150μmol/L and action time of 24 h of 6-OHDA were selected as the optimal experimental concentration and observation time for this study.Compared with 6-OHDA group,the activity of PC12 cells increased,the expression level of ROS and the apoptosis decreased in IGF-1+6-OHDA group.Conclusion IGF-1 pretreatment can reduce 6-OHDA induced oxidative damage and apoptosis of PC12 cells,also can increase cell activity,which can provide a potential strategy for the prevention and treatment of Parkinson’s disease.