摘要
帕金森症(Parkinsons disease,PD)是一种老年人群常见的中枢神经系统变性病,因发病率明显上升,其病因与防治策略亟待阐明.文中以多巴胺能神经元模型细胞PC12为材料,以致PD毒性物质鱼藤酮(Rotenone,Rot)处理细胞,检测原花青素(Grape seeds procyanidins,GSP)对Rot引起细胞毒性的防护作用,并探讨其机理.MTT法显示,GSP可减弱Rot对细胞生长的抑制作用,其效果具有浓度和时间依赖性;倒置显微镜观察吉姆萨染色和荧光染料Hoechst 33342染色后显示,Rot处理细胞由平铺多角形变为皱缩,突起回缩,细胞核染色质凝集,部分细胞核碎裂,呈现一定凋亡特征;GSP预处理对细胞和细胞核形态的改变有明显保护作用.DCFH-DA、Rhodamine 123染色后经流式细胞术分析及荧光显微镜观察显示,GSP抑制了Rot所致细胞内活性氧水平升高,同时明显改善Rot引起的线粒体膜电势降低;Annexin V/PI双染分析发现GSP联合处理明显抑制了Rot所致细胞凋亡.结果表明,GSP可能通过降低细胞ROS生成、恢复线粒体膜电势水平而保护细胞,抑制Rot引起的细胞凋亡,可能在防治帕金森病方面发挥作用.
Parkinson s disease(PD)is a common neurodegenerative disorder in elderly population.On account of increasing the incidence rate,its etiology and prevention strategy must be clarified.In this study,dopaminergic neuronal cell model PC12 were treated with rotenone(Rot),and the inhibitory effect of grape seeds procyanidins(GSP)on Rot cytotoxicity was investigated.MTT assay showed that GSP can attenuate the inhibitory effect of Rot on cell growth in dose-and time-dependent pattern.Under inverted microscope,Giemsa and Hoechst 33342 stained cells showed that Rot can damage cells significantly on cell morphology.Cell is changed from flat polygons to shrinkage,neurites is retracted,nuclear chromatin is agglutinated,and some nuclei are fragmented,showing a certain apoptosis characteristics.GSP pre-treatment has obvious protective activity on the changes of cell and nuclear morphology.After staining with DCFH-DA and Rhodamine 123,flow cytometry analysis and fluorescence microscopy showed that after GSP treatment,the content of reactive oxygen species(ROS)in cells treated with Rot is decreased significantly and the mitochondria membrane potential(MMPs)are increased.Annexin V/PI double staining test exhibited that GSP is reduced apoptosis by Rot.These results indicated that GSP can protect cells,inhibit the apoptosis induced by Rot in PC12 cell through down-regulating the production of ROS and restoring mitochondrial membrane.GSP may play a role in the prevention and treatment of Parkinson s disease.
作者
刘国安
吴亚楠
董欢
冉苗苗
孟凡雷
LIU Guo-an;WU Ya-nan;DONG Huan;RAN Miao-miao;MENG Fan-lei(College of Life Science,Northwest Normal University,Lanzhou 730070,Gansu,China)
出处
《西北师范大学学报（自然科学版）》
CAS
北大核心
2018年第3期57-64,共8页
Journal of Northwest Normal University(Natural Science)
基金
甘肃省自然科学基金资助项目(17JR5RA081)
