天津医药

天津医药 ›› 2018, Vol. 46 ›› Issue (9): 905-910.doi: 10.11958/20180149

• 细胞与分子生物学 •    下一篇

IGF-1对6-OHDA诱导神经元氧化损伤的保护作用

董晓光，许孝飞，马江波，瞿宝明，扈燕来，张静，李涛   

  1. 1青岛颐生健中西医结合骨伤医院脊柱外科（邮编266100）；2济南市第四人民医院神经外科；3宁夏医科大学人体解剖学教研 室；4山东大学基础医学院人体解剖学教研室
  • 收稿日期:2018-01-26 修回日期:2018-06-28 出版日期:2018-09-15 发布日期:2018-10-10
  • 通讯作者: 李涛 E-mail:litao961@163.com

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

DONG Xiao-guang, XU Xiao-fei, MA Jiang-bo, QU Bao-ming, HU Yan-lai, ZHANG Jing, LI Tao   

  1. 1 Department of Spine Surgery, Qingdao Orthopedic Hospital, Qingdao 266100, China; 2 Department of Neurosurgery,Jinan No.4 People’s Hospital; 3 Department of Anatomy, Ningxia Medical University; 4 Department of Anatomy of School of Basic Medical Sciences, Shandong University
  • Received:2018-01-26 Revised:2018-06-28 Published:2018-09-15 Online:2018-10-10
  • Contact: Tao LI E-mail:litao961@163.com

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摘要： 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、200 μmol/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（150 μmol/L处理24 h）和IGF-1+6-OHDA组（IGF-1 100 nmol/L预处理2 h，后加入150 μmol/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，150 μmol/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。

关键词: 胰岛素样生长因子Ⅰ, 羟基多巴胺类, PC12细胞, 氧化性应激, 细胞凋亡, 帕金森病, 6-羟基多巴胺

Abstract: 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 200 μmol/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 150 μmol/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.

Key words: insulin-like growth factor Ⅰ , hydroxydopamines, PC12 cells, oxidative stress, apoptosis, Parkinson disease, 6-hydroxy dopamine