丙炔苯丙胺对鱼藤酮致多巴胺能神经细胞损伤的保护机制研究  

Study of the Protection of Deprenyl for Neurotoxicity of Rotenone on Dopaminergic Neuron

作　　者：戚辰[1] 刘振国[1] 范国华[1] 陈生弟[1] 陆国强[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院神经内科,200025

出　　处：《中国临床神经科学》2006年第5期475-481,共7页Chinese Journal of Clinical Neurosciences

基　　金：上海市重点科委攻关资助项目(44119623);上海市教委发展基金(03BK24)

摘　　要：目的:探讨丙炔苯丙胺对多巴胺能神经细胞保护机制的研究。方法:用神经生长因子(NGF)将PC12细胞诱导分化成多巴胺能神经元的细胞模型,经鱼藤酮处理后给予不同浓度的丙炔苯丙胺,观察细胞形态改变,四甲基偶氮唑盐(MTT)法检测细胞活性及代谢状态,磷脂酰丝氨酸外翻法(Annexin-V)检测细胞凋亡,JC-1检测线粒体膜功能及DCFH-DA检测细胞内ROS。结果:经鱼藤酮处理24h后PC12细胞突起样结构消失,应用丙炔苯丙胺后细胞形态逐渐变大,突起也有恢复,细胞贴壁能力增强,与鱼藤酮组比较,在丙炔苯丙胺50μmol·L-1作用24h时即出现细胞活力恢复,A570值为0.39±0.01(P<0.01);Annexin-V检测凋亡细胞明显减少;JC-1检测丙炔苯丙胺处理组的线粒体膜电位较鱼藤酮处理组有明显恢复,DCFH-DA检测鱼藤酮组ROS水平升高,丙炔苯丙胺组ROS降低。结论:鱼藤酮在体外对多巴胺能神经元有毒性作用,丙炔苯丙胺能够明显减少鱼藤酮诱导的PC12细胞的死亡。其机制可能是抑制氧化应激及拮抗细胞凋亡。Aim： To study the protection of deprenyl for neurotoxicity of rotenone on dopaminergic neuron. Methods： PC12 cells differentiated by nerve growth factor as dopaminergic neurons were treated by different concentration of deprenyl after being treated by rotenone. Cell viability was assessed with MTF and cell apoptosis was detected by Annexin-V staining and flow cytometry. The function of mitochondrion membrane was detected by JC- 1 staining. The ROS was detected by DCFH-DA staining. Results： After treated with rotenone for 24 hours, the process-like of PC 12 cell was disappeared, and the cell body became bigger and process-like recovered after treated with deprenyl. Compared with control group treated with single rotenone, the cell viability recovered with deprenyl at 50 μmol·L^-1 concentration （A570 0.39 ± 0.01） （P〈0.01）. Apoptosis cells staining with Annexin-V were decreased treated by deprenyl following rotenone treatment. JC-1 staining showed the mitochondrion membrane＇s function recovered treated by deprenyl, and DCFH-DA staining also showed ROS in the cells decreased after treated by deprenyl. Conclusion： Rotenone is neurotoxic to dopaminergic neuron. Deprenyl can diminish cell death induced by rotenone by restraining oxidative stress and anti-apoptosis.

关 键 词：丙炔苯丙胺 鱼藤酮 帕金森病 凋亡 氧化应激 

分 类 号：R742.5[医药卫生—神经病学与精神病学]